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Published on 19 August 2016 on http://pubs.rsc.org | doi:10.1039/9781782622048-FP001
Medical Devices
Biological Fluid–Surface Interactions in Detection and
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RSC Detection Science Series
Editor-in-Chief:
Professor Michael Thompson, University of Toronto, Canada
Series Editors:
Dr Sub Reddy, University of Central Lancashire, Preston, UK
Professor Damien Arrigan, Curtin University, Perth, Australia
Titles in the Series:

1: Sensor Technology in Neuroscience
2: Detection Challenges in Clinical Diagnostics
3: Advanced Synthetic Materials in Detection Science
4: Principles and Practice of Analytical Techniques in Geosciences
5: Microfluidics in Detection Science: Lab-on-a-chip Technologies
6: Electrochemical Strategies in Detection Science
7: Peroxynitrite Detection in Biological Media: Challenges and Advances
8: Biological Fluid–Surface Interactions in Detection and Medical Devices
How to obtain future titles on publication:

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Biological Fluid–Surface
Interactions in Detection and
Medical Devices
Michael Thompson
University of Toronto, Ontario, Canada
Email: mikethom@chem.utoronto.ca
Christophe Blaszykowski
Email: cblaszyk@chem.utoronto.ca
Sonia Sheikh
Email: ssheikh@chem.utoronto.ca
Cesar Rodriguez-Emmenegger
RWTH Aachen University, Germany
Email: rodriguez@dwi.rwth-aachen.de
and
Andres de los Santos Pereira

The Academy of Sciences of the Czech Republic, Prague, Czech Republic
Email: santospereira@imc.cas.cz
Published on 19 August 2016 on http://pubs.rsc.org | doi:10.1039/9781782622048-FP001 View Online
RSC Detection Science Series No. 8
Print ISBN: 978-1-78262-097-6

PDF eISBN: 978-1-78262-204-8
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ISSN: 2052-3068
A catalogue record for this book is available from the British Library
r Michael Thompson, Christophe Blaszykowski, Sonia Sheikh,

Cesar Rodriguez-Emmenegger and Andres de los Santos Pereira 2017
All rights reserved
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Preface
The quantitative detection of species of biological interest in blood, serum,

urine, and cerebrospinal and interstitial fluids by biosensors has been the
subject of intense research over many years. The appeal of such analytical
devices resides in their promise to offer relative simplicity in their use, se-
lectivity/sensitivity for target analytes, low cost, and significant potential for
operation in both the point-of-care approach and the automated flow-
through station in the clinical biochemistry laboratory. However, despite
considerable effort over the years, fouling or non-specific adsorption effects
associated with sensor platforms have remained something of an intractable
problem. The presence of biological matrix components, such as proteins
and cells, at the sample–device interface can indeed lead to serious signal
interference. Although perhaps considered by some to constitute a separate
area of research, analogous surface phenomena are also important in terms
of the biocompatibility exhibited by medical devices, whether employed
in vivo or in extracorporeal applications. Indeed, there is much common
ground among research devoted to these topics from a surface physico-
chemical point of view. In the present book, the authors have attempted to
prepare a cohesive treatment of the surface chemistry involved in the
fouling, and its minimization, of biosensors and biomaterials in general.
The text is modular in character, with a modest level of deliberate overlap
in some chapters. We anticipate a readership originating from a disparate
set of research communities. The interested reader might very well wish to
zero-in on sections where there is less familiarity. For example, the work
opens with a concise look at the fouling problem and deals with the role of
interfacial physical chemistry in this respect (Chapter 1). An appraisal of
relevant methods for surface characterization is included in that section.

Biological Fluid–Surface Interactions in Detection and Medical Devices
By Michael Thompson, Christophe Blaszykowski, Sonia Sheikh, Cesar Rodriguez-Emmenegger
and Andres de los Santos Pereira
r Michael Thompson, Christophe Blaszykowski, Sonia Sheikh, Cesar Rodriguez-Emmenegger
and Andres de los Santos Pereira 2017
Published by the Royal Society of Chemistry, www.rsc.org
v
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vi Preface
Obviously, surface science specialists may well ‘skip’ this chapter all together
and proceed on to other aspects of the text. On the other hand, this topic
may be helpful from a learning perspective to the more biologically-inclined
reader. The book continues with a molecular view of the complex physical
chemistry that lies behind the interactions of proteins with surfaces,
pointing out basics but also warning of the perilous exercise that is to try and
establish general principles in this regard (Chapter 2). In addition to the
kinetics and thermodynamics of such processes, the chapter also includes a
look at the various models that have been proposed to explain the mech-
anism of protein–substrate interaction. The section is completed with an
overview of the practical concerns associated with protein adsorption upon
exposure of synthetic surfaces to the quintessential biofluid: blood. The
ensuing chapter then proceeds to a discussion on the interaction of bio-
logical cells with surfaces, which incorporates a brief description of relevant
cell types, mainly intended for the unfamiliar, surface science-focused
reader (Chapter 3). The section then moves on to a research area, which has
attracted enormous attention over recent years: that of the adhesion, growth,
guidance, and proliferation of cells on surfaces; all of which are especially
important in fields such as tissue engineering and regeneration. There is an
emphasis in the text on extra-cellular matrix proteins employed to orches-
trate and mimic natural tissue. Included here is a description of the effects
of substrate morphology, rigidity, and elasticity. The chapter concludes with
a concise treatment of the interaction of cells with nanoparticles, which is
crucial in the field(s) of clinical diagnostics and therapy. As implied above,
the interaction of biological entities with solid surfaces has ramifications for
the application of a wide variety of devices and systems used in modern
medicine. The next chapter deals with this subject, including topics such as
blood coagulation and inflammatory response, whose effects are crucial for
the successful implementation of biomedical equipment, implants, and
devices such as stents and vascular grafts (Chapter 4). The book then covers
the voluminous amount of work that has appeared in the literature on the
chemical surface modification of substrates with molecular coatings in an
attempt to minimize surface fouling from a variety of biofluids, whether
of human or animal origin (Chapter 5). This includes a vast array of
surface chemistries ranging from relatively simple, ultrathin self-assembled
monolayers to sophisticated, thicker polymer brush entities. The last section
of that chapter is dedicated to the assessment of the applicability of
said antifouling surface chemistry to biosensor technology. The final
chapter (Chapter 6) represents an attempt by the authors to coalesce and
rationalize past and current mechanistic arguments regarding the anti-
fouling properties of surfaces, especially the pivotal role played by surface
hydration through the inherent, molecular-level structuration of water at
interfaces. This quite naturally leads to a discussion of future surface
chemistry possibilities that are consonant with the application of biosensor
devices in biological fluids and substrates from a biocompatibility point
of view.
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Preface vii
Finally, we especially would like to express our deep appreciation to

all past and present members of the Thompson Biosensors Group at the
University of Toronto and the colleagues at the Institute for Macromolecular
Science, Czech Academy of Sciences, Prague, for their many valued research
contributions and unfailing support. We also wish to thank Debby Repka,
Edmund Chan, and Lisa Qiu of the University of Toronto for their dedicated
assistance in the preparation of some of the content of the manuscript.
Michael Thompson, Sonia Sheikh,

Christophe Blaszykowski, Cesar Rodriguez-Emmenegger
Foreword
The Physical Chemistry of Blood–Surface Interactions

Controlling the interactions between blood and synthetic material surfaces
remains as one of the most important challenges in the field of biomaterials
today. As a general rule, any foreign material surface that comes in contact
with blood becomes rapidly coated by adsorbed biomolecules (mostly pro-
teins), leading to the body’s ability to somehow recognize the surface as
‘non-self’ and react by inducing a thrombotic response. These natural pro-
cesses are, of course, in place to protect the body from vascular damage and
blood loss caused over the course of life’s normal daily activities. However,
these same processes, if left to themselves, lead to serious and often life-
threating consequences when material surfaces are intentionally placed in
contact with blood as part of a medical device (e.g., a cardiovascular stent) or
as a diagnostic procedure (e.g., an intravascular catheter or biosensor).
Of course, today’s surgical practice involves the use of a large number of
medical devices that are intentionally placed in contact with blood, ranging
from catheters, stents, and vascular prostheses, to extracorporeal systems
used during cardiopulmonary bypass surgery and renal dialysis. The clinical
approach to preventing material-induced thrombosis and the generation of
thromboemboli, which can lead to the failure of the device and/or severe
physiological complications, is to suppress the blood coagulation system on a
systemic level. This is typically done through the administration of various
anti-coagulant or anti-platelet drugs that systemically inhibit blood clot for-
mation. While widely accepted in clinical practice, the consequences of this
approach are to place patients at risk for adverse bleeding events due to the
suppression of one of the body’s most important protective mechanisms—i.e.,
blood coagulation. A much more desirable, but yet unattainable solution is to

viii
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Foreword ix
prevent thrombotic events from initiating locally at the material surface of the
medical device itself, while maintaining the body’s ability to use its natural
blood coagulation mechanisms to provide protection where otherwise
needed. Achievement of this capability requires the understanding of the
fundamental processes that govern the interactions of blood with material
surfaces; this is the primary focus of this book.

Blood is a very complex, heterogeneous fluid composed of a broad range of
macromolecules and cells within a buffered saline media. The most important
components of blood with respect to the function of medical devices placed in
contact with blood are the proteins in the blood (of which there are over 4500
different kinds) and platelets, which are the cellular mediators of blood co-
agulation. Within seconds after coming in contact with blood, material sur-
faces are generally covered with a tightly adsorbed layer of proteins. The
characteristics of this layer of adsorbed proteins (i.e., the types of proteins
adsorbed and their orientation, conformation, and packing arrangement)
have been shown to strongly influence the degree of stimulation of the
coagulation and complement cascades, as well as platelet activation. This
non-specifically adsorbed protein layer also leads to the fouling of biosensor
surfaces, compromising their sensitivity and specificity. It also provides con-
ditions that are favourable for bacterial adhesion and subsequent biofilm
formation, which can lead to chronic infection. The chapters presented in this
book take on the challenge of addressing important aspects of each of these
processes from a physical chemistry perspective, to provide the reader with
an understanding of the underlying scientific principles involved and the
practical consequences for various biomedical applications.
Chapter 1 sets the stage for the book by providing an overview of the im-
portance of the interface between synthetic materials and the biological sys-
tem. It addresses basic aspects of interfacial thermodynamics, the role of
surface morphology, methods used to physically and chemically characterize
surfaces, and methods used to characterize protein–surface interactions. This
understanding sets the stage for Chapter 2, which goes into greater depth re-
garding the physical chemistry of the interactions between proteins and sur-
faces, as well as the mathematical models used to characterize this behaviour.
This chapter continues on to address the other side of the coin, namely the
composition of blood and the blood coagulation system, and presents an
overview of the general response of blood when it comes in contact with for-
eign material surfaces. The perfect word used in this chapter to epitomize these
interactions is ‘complexity’: the complexity that involves the combined influ-
ences of protein structure, protein adsorption behaviour, and the subsequent
biological consequences of these dynamic processes; all of which combine to
make this an extremely challenging yet fascinating area of study.
While Chapter 2 focuses on interactions at the molecular level, Chapter 3
goes on to focus on cellular interactions with material surfaces. An overview of
cellular structure and function is first presented (intended primarily for ma-
terials scientists and engineers vs. biologists) along with their interactions with
substrates to mediate cell adhesion, growth, guidance, and proliferation.
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x Foreword
Examples of approaches are then presented using surface chemistry, morph-

ology, and elasticity, to control cellular responses for various types of medical
devices, including stents and extracorporeal systems used for cardiopulmonary
bypass and renal dialysis. This chapter concludes with sections dedicated to
the important topic of nanoparticles and their use for therapeutics and diag-
nostics (or in their combined form as ‘theranostics’). It also addresses con-

cerns regarding nanotoxicity, which represents a unique material surface issue
given the fact that nanosized particles are able to interact not only with the
external surface of cells, but with intracellular processes as well.
Chapter 4 continues to build on the foundation laid by the previous
chapters by going into more detail on the biological consequences of the
interactions between foreign material surfaces and blood, and addresses
topics including hemostasis, blood coagulation, inflammation, biofouling,
and biofilm formation. As pointed out in this chapter, one of the key con-
cepts for improving biocompatibility and biosensor sensitivity and speci-
ficity is to prevent the non-specific adsorption of proteins altogether. This
topic is then squarely addressed in Chapter 5, which presents an overview of
the state-of-the-art types of surface treatments and coatings that have been
developed and shown to provide a high level of resistance to non-specific
protein adsorption. The fundamental physical chemistry aspects governing
the prevention of protein adsorption are then further addressed in Chapter 6
with a focus on the important role of water structure at the interface. Finally,
Chapter 6 also provides a concluding overview of future considerations and
directions toward the long-range goal of providing truly blood compatible
surfaces for medical devices used for the diagnosis and treatment of disease.
As one who has spent a large part of his career studying these processes
and developing methods to physically characterize protein adsorption be-
haviour on material surfaces, and to understand how this influences platelet
response and blood compatibility, I see this book as a very welcome and
valuable addition to the biomaterials literature. The content of this book is
especially important for biomaterials scientists and engineers; including
students, postdocs, and professionals in both academic and industrial set-
tings, as well as for clinicians who wish to better understand the funda-
mental principles that scientists and engineers are trying to comprehend
and control through surface design. Unfortunately, the specific answers are
not yet available regarding how to achieve biomaterial surfaces for clinical
applications that are completely resistant to biofouling and the generation of
thrombus/thromboemboli without the systemic inhibition of coagulation
processes. However, the fundamental principles presented in these chapters
do provide the foundation that is needed to eventually bring the dream of
developing truly blood-compatible surfaces into reality.
Robert A. Latour
Clemson University
Clemson, SC USA
Contents
Chapter 1 Relevant Aspects of Surface Physical Chemistry 1

1.1 Introduction 1
1.1.1 Materials and Biological Systems – The
Biomaterials Interface 2
1.1.2 The Problem of Fouling 3
1.2 Interfaces and Their Properties 3
1.2.1 Definition and Thermodynamic Aspects 4
1.2.2 Surface Tension and Surface Free Energy 7
1.2.3 Wetting: Contact Angle and Capillarity 9
1.2.4 Surface Chemistry and Functionality 13
1.2.5 Mesoscopic Forces Acting on Surfaces 16
1.2.6 Surface Morphology 20
1.3 Surface Characterization Methods 24
1.3.1 Surface Chemical Characterization
Techniques 25
1.3.2 Contact Angle Techniques 29
1.3.3 Surface Structure and Morphology 31
1.3.4 Surface–Biomolecule Interactions 37
References 45
Chapter 2 Protein Adsorption on Surfaces: Understanding the

Complex Nature of a Common Phenomenon 47
2.1 Protein Adsorption on Surfaces: a Brief Overview
of a Critical Issue 47

xi
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xii Contents
2.2 Structural Complexity and Diversity of Proteins 48

2.3 Protein–Surface Interaction 52
2.3.1 A Glimpse into Protein Adsorption 52
2.3.2 Mechanism of Protein Adsorption: a
Molecular Perspective 54
2.3.3 Kinetics, Thermodynamics, and Dehydration 56
2.3.4 General Governing Principles: a Tale of
Caution 58
2.3.5 Protein Adsorption Models 59
2.4 The Case of Blood and its Plasma/Serum
Derivatives 65
2.4.1 Blood: the Proteinaceous Biofluid of Life 65
2.4.2 Deleterious Consequences of Blood-artificial
Surface Interaction in the Biomedical and
Bioanalytical Fields: an Overview 69
2.5 Conclusion 74
References 75
Chapter 3 Interaction of Cells and Tissue with Substrate Surfaces 81

3.1 Cells and Substrate Physicochemical Properties 81
3.2 The Eukaryotic Cell and Extracellular Matrix 83
3.3 Cells 88
3.3.1 Cells in Blood 88
3.3.2 The Neuron 94
3.3.3 Some Cells Used in Model Substrate Studies 96
3.4 Interaction of Cells with Substrates: Adhesion,
Growth, Guidance, and Proliferation 98
3.4.1 Bare and Treated Polymer and Metal
Substrates 99
3.4.2 Polypeptide Coatings 110
3.4.3 Extracellular Matrix Proteins and Derived
Peptides 111
3.4.4 Substrate Morphology 118
3.4.5 Substrate Rigidity and Elasticity 121
3.5 Nanoparticles, Theranostics, and Nanotoxicology 123
References 130
Chapter 4 Biological Consequences of the Blood–Surface

Interaction 136
4.1 Biological Response to Biomaterials – General
Concepts 136
4.1.1 Protein Fouling: the Initial Step 138
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Contents xiii
4.1.2Blood Coagulation and Blood–Material

Interactions 139
4.1.3 Inflammatory Reactions and the Foreign
Body Response 149
4.1.4 Complement System 152

4.1.5 Infection and Biofilm Formation 155
4.1.6 Effect of Flow Regime in Blood–Surface
Interactions 158
4.1.7 Tumorigenesis 159
4.2 Blood-Contacting Devices 159
4.2.1 Vascular Stents 159
4.2.2 Vascular Grafts 163
4.2.3 Prosthetic Heart Valves 165
4.2.4 Extracorporeal Blood Circulation 168
4.2.5 Fouling and Label-free Biosensing 170
References 177
Chapter 5 Antifouling Surface Chemistries to Minimize Signal

Interference from Biological Matrices in Biosensor
Technology 184
5.1 The Issue of Signal Interference from the Biological
Matrix in Biosensor Technology 184
5.2 State-of-the-art Antifouling Surface Chemistries 190
5.2.1 Against Blood Plasma/Serum 190
5.2.2 Against Other Human or Animal Biofluids 224
5.2.3 At-a-glance Summary 231
5.3 Implementation in Biosensor Technology 231
5.3.1 Biofunctionalization of Peptide SAMs 234
5.3.2 Biofunctionalization of Polyelectrolyte Films 235
5.3.3 Biofunctionalization of PLL-g-PEG Films 236
5.3.4 Biofunctionalization of Ionic Liquid SAMs 237
5.3.5 Biofunctionalization of Polysaccharidic
Coatings 238
5.3.6 Biofunctionalization of Polymer Brushes 240
5.4 Conclusion and Outlook 253
References and Notes 254
Chapter 6 Prevention of Deleterious Biofluid–Surface Interactions

in Detection and Medical Devices: A Look into the Future 266
6.1 On the Mechanism of Surface Resistance to Protein
Adsorption: Schools of Thought and Rationalization 266
6.1.1 Physical View 267
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xiv Contents
6.1.2 Chemical View: the Pivotal Cloaking Role of

Water 267
6.1.3 Surface Hydration and Antifouling:
A Rationalization of their Connection
through both Empirical and Computational

Evidence 269
6.2 Ultrathin Surface Chemistry for Biocompatibility
Enhancement 272
6.3 Rational Design of Biocompatible Materials 274
6.4 The Future in Practice 276
6.4.1 Antifouling Surface Chemistry: Coating vs.
Biosample Nature/Source 276
6.4.2 Implants: Tissue Healing and Bacterial
Infection 277
6.4.3 Bioanalysis and the Universalization of
Biosensor Technology 277
References 278
Subject Index 281

Published on 19 August 2016 on http://pubs.rsc.org | doi:10.1039/9781782622048-00001
CHAPTER 1
Relevant Aspects of Surface

Physical Chemistry
1.1 Introduction
Advances in biosciences, the discovery of new drugs, the development of
novel clinical diagnostic methods, and improvements in food safety rely
heavily on modern analytical techniques. These technologies provide sci-
entists with the ability not only to detect analytes, biomarkers, or specific
pathogens, but also to quantify biochemical and biophysical interactions
among biomolecules and cells. The sensitive monitoring of biomarkers
allows the early detection of disease before its progression, opening the
possibility of treatments with the highest probability of success.
Generally, the methods used in routine biochemical analyses are based on
immunoassays and require the use of labeled antibodies as reagents.
A further step, such as an enzymatic reaction, is usually necessary to produce
a physical signal that can be read. However, affinity biosensors are able to
quantify the analyte of interest without using additional reagents. They rely
on a transducer element that can directly translate the concentration of
analyte in its immediate vicinity into a physical signal. The transducer
surface must be equipped with appropriate biorecognition elements that can
bind the analyte. The advantages of label-free measurements include rapid
analysis times, real-time monitoring of concentrations, the possibility of
measuring the binding kinetics, and ease of automation and miniaturiza-
tion. Automation and miniaturization are critical, as they offer the possi-
bility of measuring a large number of analytes in parallel, with high
throughput. Such detection methods rely on the conjugation of bioreceptors.
Their continuous development depends both on advances in the design of

1
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2 Chapter 1
physical sensors and the implementation of novel sensing principles, and on

the precise control of the interactions between these artificial devices and
the biological fluids/environments in which they are meant to operate.
1.1.1 Materials and Biological Systems – The Biomaterials

Interface
An effective affinity biosensor must be able to concentrate the analyte from
the solution on its surface without eliciting non-specific interactions with
other components present in the medium. While the transducer is able to
generate a readable signal due to the presence of the analyte, it generally
cannot discriminate between changes arising from the analyte concentration
and those arising from the concentrations of interfering substances. The
biological fluids of interest for biosensing are highly complex matrices, such
as blood plasma and serum, which will be discussed in depth in later
chapters. They contain an enormous and varied range of potential
interferents.
The interactions of biosensors with biological fluids are critical for de-
termining their performance; nonetheless, many applications are affected by
such interactions. In fact, the whole category of biomaterials can be defined
as materials (other than food and drugs) that are engineered to be used,
either independently or as part of a larger system, in human or veterinary
medicine for diagnostic or therapeutic procedures involving continuous
contact with biological fluids. It is clear from this definition that affinity
biosensor surfaces fall within this category as they can perform their func-
tion only while they are in contact with the biological matrix in which the
analyte is present. Moreover, bioimplants and therapeutic devices which
come into contact with biological fluids are subject to the same phenomena.
Even outside the field of medical practice, biological and biophysical
research is constantly faced with events that occur when artificial and bio-
logical systems come into contact.
For a biomaterial to support the interactions at the interface required for
each application, the surface must be functionalized with appropriate
bioreceptors. These molecules can bind a specific target in solution or, more
generally, perform or promote a specific function. Examples include, but are
not limited to, antibodies, enzymes, aptamers, oligonucleotides, receptor
proteins, molecularly imprinted polymers (MIPs), peptide motifs, and even
whole cells. However, they need to be immobilized in such a way as to ensure
a sufficient quantity, homogeneous distribution, appropriate spatial orien-
tation, and preservation of the bioreceptor structure and conformation.
Particular applications may impose additional requirements, such as site
specificity on the bioreceptor molecule or spatial selectivity to create pat-
terns on the surface.
However, upon contact of artificial materials with complex biological
media, components of the fluid may be spontaneously deposited on the
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Relevant Aspects of Surface Physical Chemistry 3
surface. This non-specific adsorption, called fouling, can dominate the

biomaterial–biological fluid interaction, ultimately impairing the perform-
ance of a device.
1.1.2 The Problem of Fouling

The biological components most frequently responsible for fouling on
biomaterials are proteins. Their complex structures contain regions with
varying hydrophilicity and hydrophobicity, and also charges that can
mediate interactions with surfaces via multiple mechanisms. Chapter 2 will
provide an in-depth discussion of proteins and the origins of fouling. The
deposition of a layer of protein on a biomaterial surface immediately and
irreversibly alters its properties. This can affect biosensors in two ways. First,
the presence of the fouling layer can occlude the immobilized biorecognition
elements, preventing the specific binding of the analyte from taking place.
Second, label-free affinity biosensors function by directly detecting the
analyte on the basis of its intrinsic properties (e.g., mass, dielectric permit-
tivity, and electrochemical redox properties), so the properties involved in
fouling can themselves give rise to a detection signal. Since the attachment
is non-specific and does not depend on the presence of the biorecognition
elements, a signal produced by fouling can be orders of magnitude stronger
than the signal related to the specific analyte. This source of interference can
render the sensor inoperable, especially for media containing high con-
centrations of protein in solution. This is why biosensing in these media is
considered particularly challenging.
A layer of deposited protein resulting from fouling has critical con-
sequences for implantable and therapeutic devices. The proteins on the
surface mediate the subsequent in vivo response, giving cues for an immune
response. This leads to foreign body reactions and the encapsulation or
rejection of implants. When these devices come into contact with blood,
thrombogenesis is triggered, causing potentially deadly embolisms. Protein
fouling also mediates the attachment of cells and bacteria, with an increased
risk of infection.
The next section of this chapter will discuss the properties of surfaces and
the mesoscopic forces that act at the nanoscale level in terms of their rele-
vance to biosensors and biointerfaces. The last section of the chapter will
describe the experimental methods used to characterize biomaterial surfaces
and their interactions with proteins.
1.2 Interfaces and Their Properties

The interfaces between biological media and biomedical devices have a
crucial effect on their behavior and ultimately on their performance. Ac-
cordingly, the properties that describe interfaces will now be considered.
The first step is to discuss interfaces in broad terms, focusing on those
composed of solid surfaces in contact with aqueous media. It should be
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4 Chapter 1
noted that the general definitions are also applicable to the interfaces
formed between two immiscible liquids, a gas and a liquid, or two solid
phases. Emphasis will be placed on the qualitative rather than the quanti-
tative aspects. For a detailed treatment, including thermodynamic deriv-
ations of the relationships, the reader is referred to specialized texts.1–3
Precise definitions of the main interfacial properties are imperative since

they are used to characterize the behavior of materials. Understanding these
properties is essential for describing the biophysical phenomena that
determine the performance of biosensing and medical devices, the focus of
the remaining chapters of this book.
1.2.1 Definition and Thermodynamic Aspects

The term interface refers to the boundary region between two material
phases. Imagining an observer moving from one phase across the interface
into the second phase makes it possible to see that the transition of prop-
erties takes places across a certain physical volume and is not infinitely
sharp. The interface is defined as the region in which this transition takes
place. Not only does the presence of an interface imply that the two phases
coming into contact have different bulk properties, but also that new
properties emerge which are associated exclusively with the boundary
region. Consider the example of the interface formed when a droplet of oil is
suspended in water. For certain properties such as the refractive index or
viscosity, as the observer moves from inside the droplet across the interface
and into the water phase, the change will be monotonic. However, this may
not be the case for all properties of the bulk phases.
To illustrate the concept, consider a detergent, e.g., sodium dodecyl sul-
fate (Figure 1.1a), dissolved in the water phase. The chemical structure of a
detergent comprises two distinct regions: each molecule is composed of a
polar head group capable of interacting strongly with water, and a non-polar
tail with a tendency to associate with oil molecules. Its amphiphilic char-
acter causes the detergent to accumulate at the interface, each molecule
Figure 1.1 Example of a surfactant at the oil–water interface. (a) Chemical structure
of sodium dodecyl sulfate and (b) schematic representation of its con-
centration profile at the interface.
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oriented with its head group towards the water phase and its non-polar tail
towards the oil phase. If the detergent concentration is the property of
interest (Figure 1.1b), there will no longer be a monotonic trend leading
from the bulk concentration of the first phase to the bulk concentration of
the second. The concentration will have a maximum at the interface.
The many forces and interaction potentials that play a role at interfaces
will be covered later in this chapter. The appearance of specific surface- and
interface-related phenomena is of special importance in systems in which
the ratio of surface area to volume is especially high, such as colloids and
nanoparticle dispersions. The macroscopically observable properties of
these systems, including their stability and rheology, are in fact determined
by interfacial phenomena.
Now consider a system composed of two homogeneous immiscible phases
a and b (of any type) and their interface, denoted by s. From the above
discussion, it is clear that the interface exhibits physicochemical properties
that are distinct from those of either phase. Therefore, to describe the
physicochemical properties of the system as a whole, the properties of the
interface must be considered separately. For example, the internal energy
can be expressed as
U¼UaþUbþUs (1.1)
where U and U represent the internal energy of the two phases, and U s
a b
represents the internal energy of the interface region. Analogous equations

can be formulated relating other extensive properties of the system to their
counterparts in each of the phases and the interface. In particular, for each
component i, the number of moles Ni can be expressed as
Ni ¼ N ai þ N bi þ N si (1.2)
While it is obvious that the total number of moles of each component is the
sum of the numbers in the different regions, it is important to note that the
interface also contributes to the equation. In principle, these intuitive re-
lations should make it possible to determine the contributions of the
interface to the properties of the whole system simply by considering how
the properties vary across different regions and by considering the volume of
each phase V a and V b. In the bulk regions, the internal energy of the phases
is given by
U a/b ¼ V a/bu a/b (1.3)
where u a/b is the specific internal energy of each of the phases. Similarly, in a
homogeneous phase, the number of moles of each component is related to
the volume of the phase by the concentration ci
N a/b
i ¼V
a/b a/b
ci (1.4)
It follows that the number of moles in the interface is given by
N si ¼ Ni V ac ai V bc bi (1.5)
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6 Chapter 1
Figure 1.2 Variation of intensive material properties in the interfacial region. The
property can represent density, viscosity, refractive index, etc.
However, as mentioned previously, the change in a property that takes place

between the bulk regions of the two phases occurs over a narrow but not
infinitely thin region. Accordingly, the values obtained for the quantities
associated with the interface will depend on where the interface is defined to
be. Even knowing that the sum of the volumes of the two phases equals
the total volume does not determine exactly where one phase begins and the
other ends. Thus, although the total volume V of the system is known, the
individual volumes V a and V b of the phases are not. It is still necessary to
pinpoint the location of the interface (Figure 1.2).
Following the convention introduced by Gibbs, all the properties of
the interface can be assigned to an infinitely thin plane, and the boundary
between the phases can be defined so that for one major component, the
excess of moles at the interface is zero. For example, let the phase a be an
aqueous solution, as is the case for biological fluids. Then the interface can
be taken to be the dividing plane between the bulk phases such that the
excess number of moles of water at the interface is zero:
N sw ¼ 0 (1.6)
Knowing the concentrations caw and cbw of water in both bulk phases, the
above condition makes it possible to define the volume of the aqueous so-
lution (phase a) using eqn (1.5). This fixes the position of the delimiting
plane. Intuitively, caw4cbw because a is an aqueous phase immiscible with b.
From eqn (1.5) and (1.6), and the condition that the total volume is the sum
of the volumes of a and b, it is clear that there is only one possible value of
V a, and therefore one value of Vb, since the total volume of the system is
fixed and V ¼ V a þ V b. At this stage, it is possible to calculate the contri-
bution of the interface to the thermodynamic internal energy of the system:
U s ¼ U V aua V bu b (1.7)
For the purpose of thermodynamic analysis, the interface can be con-
sidered to be an infinitely thin surface with area A, which can be determined
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according to the Gibbs criterion. It is useful to define the intensive interface

properties, such as the specific interface internal energy:
Us
us ¼ (1:8)
A
1.2.2 Surface Tension and Surface Free Energy

For homogeneous systems, the enthalpy is defined as H U þ pV, and it is a
state function. For many systems and processes, especially in chemistry and
biology, enthalpy is the preferred quantity for calculating energy balances
because a change in enthalpy at constant pressure equals the heat input into
the system plus the work output from the system other than pressure–
volume expansion work. However, once an interface is introduced into the
system, the mechanical work must include not only the pressure–volume
work pDV, but also the work required to enlarge or shrink the interface. This
change can be associated with the difference DA in the surface area of the
interface. The following paragraphs describe how this work can be
incorporated.
The key concept that must be considered at this stage is that the creation
of an interface or the extension of an existing interface requires the input of
work. Therefore, there is a certain energy associated with the existence of an
interface that depends on its dimensions.
Consider a liquid supported by a rectangular frame on three sides with a
movable slider of length l on the fourth side, under constant pressure and
temperature (Figure 1.3). Pulling the slider a distance d will increase the area
Figure 1.3 Interfacial tension. A layer of liquid is held by a frame on three sides,
while the fourth side is able to slide. As it is pulled under constant force,
the surface area of the layer of liquid is extended on both sides.
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8 Chapter 1
of the liquid interface by DA ¼ 2ld The factor of 2 appears because the new
interface area is created on both sides of the liquid layer. Such movement
will require a force F to be applied, which is constant throughout the pulling
process. The absolute value of the force exerted on the liquid to increase its
area is given by
|F| ¼ 2l g (1.9)
The proportionality factor g is known as the surface tension of the liquid,

and it has units of force divided by distance (N m1). The work required for
this process is proportional to the increase in the area of the liquid. It is
given by
W ¼ g DA (1.10)
Since the process occurs under constant pressure and temperature, this
work equals the increase in the Gibbs free energy of the system. As a result, g
is also the increase in the surface free energy:

@G
g¼ (1:11)
@A T;P
The excess free energy of a surface is the result of the loss or rupture of
chemical bonds and/or attractive van der Waals and electrostatic inter-
actions of the molecules at the interface; these losses arise because there are
fewer neighboring molecules than in the bulk phase. Therefore, liquids in
air tend to minimize their surface area. As an example, a droplet of water in
space not subject to gravity tends to have a spherical form since this shape
minimizes the surface area to volume ratio.
In a solid, the creation or enlargement of the surface area is also accom-
panied by an increase in free energy. Likewise, the increase in the free
energy of the system can be expressed by defining the surface free energy as
in eqn (1.11).
The concepts of surface tension and surface free energy are essential for
the study of processes in which new interfaces are created. Wetting and
adhesion are two important examples of processes in which changes in area
are determined by the interplay of surface free energies. Wetting will be
explained in more detail in the following section.
Typically, solids are classified into high energy and low energy solids ac-
cording to the type of bond that holds the molecules together. Solids of the
first type are composed of networks of covalent or ionic bonds, which cannot
be broken without a large amount of energy. In contrast, in low energy
solids, individual molecules are attracted to each other due to much weaker
van der Waals interactions or hydrogen bonding. The energy of these bonds,
which must be overcome to form a new solid surface, determines the surface
free energy of a solid. In practice, because of energy dissipation and plastic
deformation during the process of breaking, the energy input required to
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split a solid and create two new surfaces can be much greater than the
energy of the bonds involved.
The surface tension of liquids follows an analogous trend. Alkanes,
interacting only through van der Waals dispersion forces, have very low
surface tensions. The behavior of water molecules is different; they form a
network involving hydrogen bonds, which is disrupted by the presence of an

interface. This carries a large free energy penalty, and consequently, the
surface tension of water is high.
1.2.3 Wetting: Contact Angle and Capillarity

The presence of a surface renders a contribution, which is always positive, to
the total free energy of a system. The tendency to minimize the surface free
energy is evident in the wetting phenomena observed in systems consisting
of multiple phases that come into contact, and of the resulting interfaces.
The example of the interfaces created when a droplet of liquid comes into
contact with a solid surface will be addressed in detail due to the importance
of such systems in many practical applications. Similar surface energy
arguments apply to systems consisting of immiscible liquids that come into
contact.
As a droplet touches a solid, three interfaces are created: solid–air, liquid–
air, and solid–liquid (Figure 1.4). The spreading of the droplet on the surface
increases the area of the solid–liquid interface at the expense of the solid–air
interfacial area. The tendency of a droplet of liquid to spread on the solid
surface depends on the relative interfacial free energies of the solid–air,
liquid–air, and solid–liquid interfaces. Equilibrium is reached when the
interfacial areas are such that the free energy of the system is minimal. In
this situation, the interfacial tensions can be analyzed at the point of contact
of the three phases. At equilibrium, the sum of the components of the forces
parallel to the surface must be a zero net force acting on the contact region
of all the phases:
gSL þ gLA cos(y) gSA ¼ 0 (1.12)
Figure 1.4 Contact angle at the interface of a solid and a liquid in air. Under
equilibrium conditions, the contact angle y formed is such that the
components of the interfacial tensions parallel to the surface are
balanced and Young’s equation is fulfilled.
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10 Chapter 1
where gSL, gLA, and gSA are the interfacial tensions of the solid–liquid, liquid–
air, and solid–air interfaces, respectively. The condition given by eqn (1.12),
known as Young’s equation for the contact angle, determines the spreading
of the droplet on the solid surface. The equilibrium contact angle can be
calculated from the surface free energies of the interfaces. The more ener-
getically favorable the spreading of the droplet (i.e., increasing the solid–
liquid area at the cost of decreasing the solid–air interface area), the lower
the contact angle and the larger cos(y). In the extreme case, a contact angle
of zero amounts to complete spreading of the liquid on the solid surface,
and the solid is said to be fully wettable by the liquid. This can occur, for
example, with low energy liquids or high energy solids. At the other end of
the wettability scale, very high contact angles, e.g., greater than 901, can be
observed for water on smooth fluorinated surfaces such as PTFE, which has
extremely low surface energy. The most important liquids for applications in
the biological sciences are water and its solutions. Surfaces that are easily
wettable by water, i.e., those having low contact angles, are called hydro-
philic; materials that repel water and have high contact angles are called
hydrophobic. The affinity of a surface for water is critically important for
determining its interactions with biological media and components, as will
be discussed in subsequent chapters of this book. The most important
physicochemical features influencing the hydrophilicity/hydrophobicity of a
material surface will be addressed later in this chapter in the context of
surface chemistry and surface chemical functionality.
The contact angle of water or other liquids on a surface can be a valuable
source of information about the surface free energy and hence the surface
chemistry.4 However, this parameter cannot be obtained directly from
Young’s equation since it contains two unknown variables, gSL and gSA (the
surface tension gLA of a large range of liquids at different temperatures can
be found in the literature). A quantity commonly used to approximate the
energy of a surface is the critical surface tension, which can be obtained by
constructing a Zisman plot. The critical surface tension is defined as the
maximum surface tension of a liquid that will fully wet the surface. A liquid
having a surface tension higher than the critical surface tension of a solid
will not spread completely and instead will form a finite contact angle.
A Zisman plot, based on measurements of the contact angles between
a surface and various liquids having different surface tensions, graphs
cos(y) vs. gLA. The critical surface tension is obtained as the intercept at
cos(y) ¼ 1 of the extrapolated line of the experimental points. Figure 1.5
shows the calculation of the critical surface energy for PTFE.5
However, several objections have been raised about the relationship of the
critical surface tension to the performance of a surface in biomedical ap-
plications. To begin with, critical surface tension values are usually found for
low surface tension liquids, such as alkanes and other organic solvents, to
avoid the swelling of the solid by the liquid. However, it is the interactions
with an aqueous environment that are critical for bioimplant and biosensing
applications. The behavior of a surface in contact with air or low surface
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0.7
0.8
cos(θ)
Critical surface
tension
0.9
1.0
18 20 22 24 26 28
Liquid surface tension (mN m-1)
Figure 1.5 Calculation of the critical surface tension of a solid by the Zisman
method. The contact angles on the surface of PTFE were measured
with a series of di(n-alkyl)ethers. The intersect of the linear fit at
cos(y) ¼ 1 represents the critical surface tension.
(Prepared from data from Table I in ref. 5.)
tension liquids may not be a good predictor of its behavior at an interface

with water.6 The quantification of the energy of interaction with non-polar
liquids will yield only the dispersive component (i.e., London forces; see
below) of the surface free energy. Another problem is that Zisman plots are
often non-linear. In addition, while the Zisman plots used to calculate the
critical tension may have a reasonably good fit for a series of liquids within
the same family of compounds (e.g., a series of alkanes, a series of halo-
carbons, or a series of alkyl ethers), the values obtained for different families
can vary due to the interactions between the liquids and the surface.
The wetting of a smooth surface by a droplet of liquid is a dynamic pro-
cess, and usually a different contact angle is observed for a droplet whose
volume is increasing than for a droplet whose volume is decreasing. When a
droplet of liquid is placed on a surface and its volume is slowly increased by
the careful addition of liquid (for example, through a needle connected to a
syringe pump), the contact area begins to expand, and the droplet forms a
characteristic advancing contact angle. If the process is reversed and liquid
is slowly drawn out of the droplet, thereby decreasing its volume, initially the
contact area remains unchanged, and the droplet will be pinned while the
contact angle decreases. If liquid continues to be drawn out of the droplet,
reducing its volume, at some point, the contact area will start to decrease,
and the droplet forms a characteristic receding contact angle. The equi-
librium contact angle, as calculated from the interfacial energies and
Young’s equation, lies between the advancing and receding contact angles.
The difference between the advancing and receding contact angles is called
contact angle hysteresis, for which there are various causes. While hysteresis
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luck may turn. You’ll not always be what you are now. Suppose you
have a bad illness?”
“I’m thinking of his side of the case.” The tone verged upon
sternness.
“You have quite enough to do to think of your own. Don’t throw
chances away. I have had forty years’ experience of a very hard
profession, and even you top sawyers are on very thin ice. And
remember, the cards never forgive. Girls who have a lone hand to
play, mustn’t hold their heads too high. If they do they’ll live to regret
it. And you mustn’t think these swells can’t box their own corner.
They’ve nothing to learn in looking after Number One. A girl of your
sort is quite equal to any of these drawing-room noodles and Mr.
Dinneford knows that better than I do.”
“But that’s impossible. I can never be as they are.”
“You needn’t let that worry you. A lot of stuck-up dunces that all
the world kow-tows to!”
“It isn’t that I think they are nicer or cleverer or wiser than other
people. But they are born to certain things, they have been bred to
them for generations, and it surely stands to reason that they are
better at their own game than a mere outsider can hope to be.”
“Fiddle-de-dee!” said Mrs. Wren. “I hope you are not such a
goose as to take swelldom at its own valuation. It’s all a bluff, my
dear. Your humble servant, Jane Wren, could have been as good a
duchess as the best of ’em if she had been given the chance. I don’t
want to be fulsome, my dear, but I’ll back a girl of your brains against
Lady Agatha Fitzboodle or any other titled snob.”
“But I don’t want to be pitted against anybody!”
“That’s nonsense.” Mrs. Wren shook a worldly-wise head. “As for
being an outsider, a girl can’t be more than a lady just as a man can’t
be more than a gentleman. And if you are a lady and have always
gone straight you needn’t fear comparison with the highest in the
land.”
Mary shook a head of sadness and perplexity.
“Somehow it doesn’t seem right to mix things in that way,” she
said.
“It’s the only way that keeps ’em going,” said Mrs. Wren
scornfully. “And well they know it. At least nature knows it. Look at
Wrexham! Do you mean to say that his inbred strain wouldn’t be
improved by Milly? And it’s the same with you and Mr. Dinneford. It’s
Nature at the back of it all. It’s the call of the blood. If these old
families keep on intermarrying long enough dry rot sets in.”
Mary stood a picture of woe.
“You odd creature!” said Mrs. Wren. “I’ve never met a girl with
such ideas as yours. I really believe you are quite as narrow and as
prejudiced as Lady Agatha Fitzboodle. To hear you talk one would
think you believed rank to be a really important matter.”
Incredulous eyes were opened upon the voluble dame.
“Of course it is.” But the girl’s solemnity was a little too much.
“My dear!” A gust of ribald laughter overwhelmed her. “Hasn’t it
ever struck you that the so-called aristocracy racket is all a bluff?”
“Surely, it can’t be.” The tone was genuine dismay.
“Every word of it, my dear. There’s only one thing behind it and
that’s money. If Wrexham ever sticks a coronet on the head of my
Milly and robes her in ermine she’ll be the equal of any in the land,
just as old Bill Brown who was in the last birthday honors is as good
a peer as the best of ’em now that his soap business has brought
him into Park Lane. I knew Bill when he hadn’t a bob. It’s just a
matter of L.S.D. As for the frills, they are all my eye and Elizabeth
Martin. When my Milly gets among them, it won’t take her a week to
learn all their tricks. They are just so many performing dogs.”
“You don’t understand, you don’t understand!” The tone was
tragic.
VIII
A night’s reflection convinced the girl that there was only one
thing to be done. The engagement must end. But as she soon found,
it was easier to make the resolve than to carry it out. To begin with, it
was terribly irksome, in present circumstances, to give effect to her
decision and to back it with reasons.
Her début in the Row had been so successful that a ride had
been arranged for the next morning. But it was spoiled completely by
the specter now haunting her. In what terms could she tell him that
she had changed her mind? How could she defend a proceeding so
unwarrantable?
It was not until later in the day, when they took a stroll under the
trees in the Park, that she forced herself to grasp the nettle boldly.
Jack, as she had foreseen, was immeasurably astonished. He
called, at once, for her reasons. And they were terribly difficult to put
into words. At last she was driven back upon the cardinal fact that he
had concealed his true position.
He repudiated the charge indignantly. In the first place, he had
taken it for granted that she knew his position, in the second, he
always made a point of leaving it as much as possible outside his
calculations.
“But isn’t that just what one oughtn’t to do?” she said, as they
took possession of a couple of vacant chairs.
“To me the whole thing’s absurd,” was the rejoinder. “It’s only by
the merest fluke that I have to succeed to the title, and I find it quite
impossible to feel about things as Bridport House does. The whole
business is a great bore, and if a way out could be found I’d much
rather stay as I am.”
“But isn’t that just a wee bit selfish, my dear—if you don’t think
me a prig?”
“If you are quite out of sympathy with an antediluvian system, if
you disbelieve in it, if you hate it in the marrow of your bones,
where’s the virtue in sacrificing yourself in order to maintain it?”
“Noblesse oblige!”
“Yes, but does it? A dukedom, in my view, is just an outworn
convention, a survival of a darker age.”
“It stands for something.”
“What does it stand for?—that’s the point. There’s no damned
merit about it, you know. Any fool can be a duke, and they mostly
are.”
Mary, if a little amused, was more than a little shocked.
“I’m sure it’s not right to think that,” she declared stoutly. “I would
say myself, although one oughtn’t to have a say on the subject, that
it’s the duty of your sort of people to keep things going.”
“They are not my sort of people. I was pitchforked among them.
And if you don’t believe in them and the things it is their duty to keep
going what becomes of your theory, Miss Scrupulous?”
“But that’s Socialism,” said Mary with solemn eyes.
“No, it’s the common sense of the matter. All this centralization of
power in the hands of a few hard-shells like my Uncle Albert—he’s
not my uncle really—is very bad for the State. He owns one-fifth of
Scotland, and the only things he ever really takes seriously are his
meals and his health.”
“He stands for something all the same.”
The young man laughed outright.
“I know I’m a prig.” The blushing candor disarmed him. “But if one
has a great bump of reverence I suppose one can’t help
exaggerating one’s feelings a little.”
“I suppose not,” laughed the young man. And then there was a
pause. “By jove,” he said at the end of it, “you’d be the last word in
duchesses.”
“You won’t get Bridport House to think so.”
“So much the worse for Bridport House. Of course, I admit it has
other views for me. But the trouble is, as always in these close
corporations, they haven’t the art of seeing things as they are.”
Mary shook a troubled head, but the argument seemed to find its
way home.
“The truth of the matter is,” he suddenly declared, “you are afraid
of Bridport House.”
Without shame she confessed that Bridport House was bound to
be very hostile, and was there not every reason for such an attitude?
Jack, however, would not yield an inch upon that count, or on any
other if it came to that. He was a primitive creature in whom the call
of the blood was paramount. Moreover, he was a very tenacious
fellow. And these arguments of hers, strongly urged and boldly
stated, did not affect his point of view. The ban of Fortune was purely
artificial, it could not be defended. She was fain, therefore, to carry
the war to the enemy’s country. But if she gently hinted a change of
egotism he countered it astutely with the subtler one of
sentimentalism. Each confessed the other partially right, but so far
from clearing the air it seemed to make the whole matter more
complex. The upshot was that he called upon her to find a valid
reason, otherwise he refused point-blank to give her up.
“Just think,” he said, tracing her name on the gravel with a
walking-stick, “how hollow the whole business is. How many of Uncle
Albert’s ‘push’ have married American wives without a question?
And why do they, when they wouldn’t think of giving English girls of
the same class an equal chance? In the first place, for the sake of
the dollars, in the second, because it is so easy for them to shed
their relations and forget their origin.”
But so wide was the gulf between their points of view that mere
argument could not hope to bridge it. If she was in grim earnest, so
was he; moreover she had entered into a compact he was
determined she should fulfill. Before consenting to release her she
would have to show very good cause at any rate.
Suddenly, in the give-and-take of conflict, Laxton came into her
mind. The memory of Beaconsfield Villas, the whimsical creatures of
another orbit, and the childhood which now seemed ages away, fired
her with a new idea. She would take him to see the humble people
among whom she had been brought up.
CHAPTER V
ALARUMS AND EXCURSIONS
I
The flight of time had affected Beaconsfield Villas surprisingly little.
Laxton itself had deferred to Anno Domini in many subtle ways; it
had its electric trams and motor-buses, and the suburb had doubled
in size, but no epoch-making changes were visible in the front sitting-
room of Number Five. In that homely interior the cosmic march and
profluence was simply revealed by a gramophone, the gift of Mary,
on the top of the sewing machine in the corner, and by the accession
to the walls of lithograph portraits of the son and grandson of the
august lady who still held pride of place over the chimney-piece.
The afternoon was stifling even for South London in the middle of
June. And Joseph Kelly, who had attained the rank of sergeant in the
Metropolitan Police Force, not having to go on duty until six o’clock
that evening, was seated coatless and solemn, spectacles on nose,
smoking a well-colored clay and reading the Daily Mail. At the level
of his eyes, in portentous type was, “Laxton Bye-Election. A Sharp
Contest. New Home Secretary’s Chances.” Joe was a shade stouter
than of yore, his face was even redder, a thinning thatch had turned
gray, but in all essentials the man himself was still the genial
cockney of one-and-twenty years ago.
The outer door of the sitting-room, which was next the street, was
wide open to invite the air. But ever and again there rose such a
fierce medley of noises from a mysterious cause a little distance off,
that at last Joe got up from his chair, and waddling across the room
in a pair of worn list slippers, banged the door against the sounds
from the street which had the power to annoy him considerably.
Hardly had Joe shuffled back to his chair and his newspaper
when the door was flung open again and an excited urchin thrust a
tousled head into the room.
“‘Vote for Maclean an’ a free breakfast-table’!”
The law in the person of Sergeant Kelly rose from its chair
majestically.
“If you ain’t off—my word!”
Headlong flight of the urchin. Joe closed the door with violence
and sat down again. But the incident had unsettled him. He seemed
unable to fix his mind on the newspaper. And the noises in the street
waxed ever louder. Now they took the form of cheers and counter
cheers, now of hoots, cat-calls and shouts of derision. At last the
tumult rose to such a pitch that it drew Eliza from an inner room.
The years had changed her rather more than her husband. But
she was still the active, capable, bustling housewife, with a keen eye
for the world and all that was passing in it.
“They are making noise enough to wake the dead.” Eliza looked
eagerly through the window.
“I wish that durned Scotchman hadn’t set his committee-room
plumb oppersite Number Five, Beaconsfield Villas,” was Joe’s sour
comment.
At that moment the all-embracing eye of a relentless housewife
swooped down upon a card lying innocently on the linoleum. It had
been flung there by the recent visitor. Eliza picked it up and read:
Vote for Maclean, thus:

Maclean X
Whitley.
On the back of the card was a portrait of Sir Dugald Maclean,

M.P.
Eliza gazed at it in astonishment mingled with awe.
“I am bound to say he is a better-favored jockey than when he
came a-courting our Harriet. Look, Joe!”
With scornful vehemence, Joe declined the invitation.
Eliza was sternly advised to tear up the card, but instead she
chose to set it on the chimney-piece. The rash act was too much for
her lord. Once more he rose from his chair, tore the card into little
pieces and flung them into a grate artistically decorated with colored
paper.
“You are jealous!” said Eliza, laughing.
“Of the likes of him! Holy smoke! But if you think we are going to
have such trash in the same room as the Marquis, you make an
error.”
The words had hardly been uttered when shouts yet more
piercing came from the street. Eliza made a hasty return to the
window.
“Come and look, Joe!” she cried breathlessly. “Here he is with his
top hat and eyeglass. He’s that dossy you wouldn’t know him. He’s
dressed up like a tailor’s dummy.”
But Joe declined to budge.
“It fairly makes me sick to think of the feller,” he said.
A little later, when the tumult in the street had died down a bit,
Joe settled himself in his chair for an afternoon nap. Eliza, duly
noting the symptoms, retired on tiptoe to another room, closing the
door after her gently. But today, alas, the skyey influences were
adverse. Joe had barely entered oblivion when a smart tap at the
street door shattered this precarious peace. With a grudge against
society he rose once more, shambled across the room and flung
open the door, half expecting to find that the urchin had returned to
torment him. A dramatic surprise was in store. On the threshold was
a creature so stylishly trim that even the blasé eye of the
Metropolitan Force was sensibly thrilled in beholding her. “A bit of
class” without a doubt, although adorned by the colors of the
People’s Candidate, and surprisingly cool in sheer defiance of the
thermometer.
“Good afternoon!” The tone of half-confidential intimacy was quite
irresistible. “May I have a little talk with you?”
“Certainly, miss.” The unconscious gallantry of an impressionable
policeman was more than equal to the occasion. “Step inside and
make yourself at home.”
When Joe came to review the incident afterwards, it seemed very
surprising that he should have yielded so easily to the impact of this
elegant miss. For instinctively he knew her business. Moreover, the
last thing he desired at that moment was to be troubled by her or by
it. But he had been taken by surprise, and in all circumstances he
would have needed ample notice to deny a lady. He had a great but
impersonal regard for a lady, as some people have for a Rembrandt
or a Corot or a Jan van Steen. And although the fact was not
important, perhaps his sense of humor was a little touched by such a
young woman taking the trouble to come and talk to such a man as
himself.
“I am here,” said the voice of the dove, as soon as its owner had
subsided gracefully upon a chair covered with horsehair, “to ask your
vote and interest for Sir Dugald Maclean, the People’s Candidate.”
The prophetic soul of Joe had told him that already. But again the
sense of humor, the fatal gift, may have intervened. Had the elegant
miss had any nous, she would have known that a sergeant of the X
Division has not a vote to bestow. In justice to the fair democrat, Joe
might have reflected that in the absence of his tunic there was
nothing to show his status. However, he didn’t trouble to do that. It
was enough for him that she was on a fool’s errand. But Joe was a
man of the world as well as a connoisseur of the human female. A
picturesque personality intrigued him. Moreover, it was working for a
cause that Joe despised from the depths of his soul. So much was
she “the real thing” that she had even turned on a melodious lisp for
his benefit; yet he had no particular wish, even under these flattering
auspices, to discuss the people and their champion. He had quite
made up his mind about both. But, the Machiavellian thought
occurred to him, here was a dangerous implement in the hands of
the foe, therefore it would be the part of wisdom to waste a little of
her time.
“‘Government of the people, by the people, for the people,’”
lisped the siren, “that, of course, as you may know, is what Sir
Dugald stands for.”
“Does he!” reflected Joe. With a roguish smile he looked the
speaker over from her expensive top to her equally expensive toe.
“You do believe in the people?” said the siren with a rather
dubious air.
“Since you ask the question, miss,” said Joe, “I am bound to say I
don’t, and never have done.”
“Not believe in the people!” It didn’t seem possible.
“If you’d seen as much of the people as I have, miss,” said Joe
grimly, “I’m thinking you’d not be quite so set up with ’em.”
The tone of conviction disconcerted the fair canvasser. Somehow
she had not expected it. In the course of her present ministrations it
was the first time she had met that point of view. Laxton’s working-
class, which for several days had been honored by her delicate
flatteries, had shown such a robust faith in itself and had purred so
responsively to her blandishments that she now took for granted that
in all circumstances it would fully share her own enthusiasm for it.
But this rubicund, coatless Briton, with eyes of half truculent humor,
was a little beyond her. Gloves were needed to handle him;
otherwise fingers of such flowerlike delicacy stood a chance of being
bruised.
“May one ask what you have against them?” lisped the people’s
champion, opening large round eyes.
“Nothing particular, miss,” said Joe urbanely. “But you ask me
whether I believe in ’em and I say I don’t. Mind you, the people are
all right in their place. I’ve not a word to say against ’em personally.
Of a Monday morning at Vine Street, when the Court has been swep’
an’ dusted and his Worship has returned from his Sunday in the
country, we always try to make ’em welcome. ‘Let ’em all come,’
that’s the motto of the Metropolitan Force. But as for believing in ’em,
that’s another story.”
This was rather baffling for the people’s champion. She was at a
loss. But her faith was sublime. This odd, crass, heavy-witted
plebeian who denied his kind was a sore problem even for the
bringer of the light. Still, she stuck to her guns gallantly.
“‘Government of the people, by the people, for the people.’”
Lisping the battle cry of Demos she returned stoutly to the charge.
Sacred formulas flowed from her lips in a stream of charming
pellucidity.
“Ah, you don’t know ’em, miss,” ejaculated Joe, at intervals.
It was a pretty joust; vicarious enthusiasm on the one side, first-
hand experience on the other. But Joe was a rock. The fair
canvasser took forth every weapon of an elegantly-furnished armory,
yet without avail.
“I don’t hold with the people, miss, not in no shape nor form.”
The tone was so final that at last a sense of defeat came upon
this Amazon. She was still seated, however, without having quite
made up her mind to the inevitable, on her grand chair in the front
sitting-room of Number Five, Beaconsfield Villas, when Fate
intervened in quite a remarkable way.
All of a sudden, there appeared on the threshold of the open door
a figure tall, fine and unheralded. It was that of Harriet Sanderson.
“Anybody at home?” she inquired gayly.
The unexpected visitor was looking very handsome and
distinguished in a well-cut black coat and skirt, and a large hat too
plain for fashion, but very far from démodé. She came into the room
with that almost proprietary air she was never without in her
intercourse with her own people. But it was about to suffer an
eclipse.
Harriet just had time to greet her brother-in-law with a happy
mingling of the bon camarade and the woman of the world, her fixed
attitude towards such an Original, whom somehow she could not
help liking and respecting, when her eyes met suddenly those of the
fair canvasser.
For a moment an intense surprise forbade either to speak. But
the people’s champion was the first to overcome the shock.
“Mrs. Sanderson!” she exclaimed.
The change in Harriet was immediate and dramatic.
“Lady Muriel!” A slight flush of a fine face accompanied the tone
of awe.
The visitor rose. And in the act of so doing an accession of great
ladyhood, almost entirely absent a few minutes ago, seemed
automatically to enter her manner.
“What a small world it is!” she laughed. “Fancy meeting you
here!”
By now the iron will of the secretly annoyed and oddly
discomposed Harriet was able to reassert itself.
“It is a small world, my lady.” The tone was a very delicate
mingling of aloofness and respect.
Brief explanations followed. These quickly culminated in the
presentation of Joe, who then became the most embarrassed of the
three. Unawares and in his shirt sleeves, he had been entertaining
an angel. And to one of Conservative views, with a profound
reverence for law, order and all established things, this seemed to
verge upon indecency. A mere “one of Scotchie’s lady canvassers”
had been magically transformed, in the twinkling of an eye, into Lady
Muriel Dinneford, the third daughter of one whom Number Five,
Beaconsfield Villas, always alluded to as “his Grace.”
II
It was the work of a few tactful minutes for Lady Muriel to effect a
discreet retirement from the scene. Yet so deeply had she been
engaged by Joe’s contumacy, and at the back of a mind which was
making the most heroic efforts to be “broad” was such a sense of
amusement, that she declared her intention of returning anon with
the People’s Candidate, if he could possibly spare a few minutes
from his multifarious duties, in order that the coup de grâce might be
given to Mr. Kelly’s dangerous heresies.
The withdrawal of the distinguished visitor across the street to the
Candidate’s committee room left a void which for a few tense
moments only wonder could fill.
It was Joe who broke the silence which, like a pall, had suddenly
descended upon the front parlor of Number Five.
“If that don’t beat Banagher,” he said. “Fancy one of the Fam’ly
taking the trouble to come a canvassin’ for Scotchie!”
Keen humor and acute annoyance contended now in the
eloquent face of Harriet.
“Pray, why shouldn’t she canvass for Sir Dugald Maclean”—the
level voice was pitched in a very quiet key—“if she really believes in
his principles?”
“How can she believe in ’em, gal?”
“Why not?”
“How can a blue blood believe in that sort of a feller?”
“Sir Dugald is a remarkably clever man. One of the cleverest men
in England, some people think.”
“That’s nothing to do with the matter. It’s character that counts.”
“There’s nothing against his character, I believe. At any rate,
Lady Muriel is going to marry him.”
The state of Joe’s feelings forbade an immediate reply. And when
reply he did, it was in a tone of scorn. Said he: “‘Government of the
people, by the people, for the people!’ Harriet, for a dead beat fool
give me a blue blood aristocrat.”
“Joe,” came the answer, with a gleam of humor and malice, “I
really think you should learn to speak of our governing class a little
more respectfully.”
This was rather hard. She ought to have realized that it was
because Joe respected them so much that he now desired to
chasten them.
“Scotchie of all people!” he muttered.
“There’s no accounting for taste, you know.” There was a sudden
flash of a very handsome pair of eyes.
“O’ course there ain’t,” said Joe, sorrowfully malicious. “You may
have forgot there was a time when Scotchie came a-courtin’ you.”
“Do you suppose I am ever likely to forget it!” said Harriet, with a
cool cynicism which took the simple Joseph completely out of his
depth.
“Well, it’s a queer world, I must say.”
“It is,” his sister-in-law agreed.
At that moment, Eliza came into the room. The visit of Harriet
was so unexpected as to take her by surprise. But the cause of it
was soon disclosed. Harriet was troubled about Mary. Ever since the
girl, against the wishes and advice of her friends, had taken what
they felt to be a fatal step, there had been a gradual drifting apart.
Harriet had kept in touch with her as well as she could, but she had
not been able to stifle her own private fears. The peril of such a
career, even when crowned by success, was in her opinion, difficult
to exaggerate. She disapproved of the friendship with the Wrens,
and had strongly opposed Mary’s living with them. But as the girl
rose in her profession, Harriet’s hold upon her grew still less. And
now at second and third hand had come news which had greatly
upset her.
With the tact for which she was famous, Harriet did not speak of
this in the presence of Joe. She accompanied Eliza to the privacy of
the best bedroom, ostensibly to “take off her things,” but really to
discuss a matter which for the past week had filled her with
misgiving.
In the meantime, Joe in the parlor set himself doggedly to
compass the nap that so far had been denied him. In spite of the
noises in the street and romantic appearance of a real live member
of the Family in his humble abode, he had just begun to doze when
the ban of Fate fell once more upon him.
From the strange welter in the amazing world outside there now
emerged a large open motor. And royally it drew up before the magic
door of Number Five. Two persons were seated in the car. One was
no less than Princess Bedalia. The other was the humblest and yet
the boldest of her adorers.
III
The idea itself had been Mary’s that they should use a fine
afternoon in motoring into Laxton, in order to see her parents.
Behind this simple plan was fell design. A week had passed since
that conversation under the trees in the Park in which she had
sought in vain for her release. But so shallow had her reasoning
appeared that Jack declined to take it seriously. He had her promise,
and he felt he had every right to hold her to it. Unless she could
show a real cause for revoking it, he was fully determined not to give
her up.
In desperation, therefore, she had hit on the expedient, a poor
and vain one, no doubt, of taking him to see those humble people
whom she called father and mother. In the course of her twenty odd
years up and down the world she had had intimations from various
side winds and divers little birds that she was an adopted child. Her
real parentage and the circumstances of her birth were an
impenetrable mystery and must always be so, no doubt, but her
feeling for the Kellys was one of true affection and perfect loyalty.
Not by word or deed had she hinted at the possession of knowledge
which had come to her from other sources.
In the circumstances of the case she now allowed herself to
imagine that a visit to her home people in their native habit as they
dwelt might help to cure Jack of his infatuation. An insight into things
and men told her that Beaconsfield Villas must be whole worlds
away from any sphere in which he had moved hitherto. Nor would he
be likely to suspect, as she was shrewdly aware, that a creature so
sophisticated as herself had risen from such humble beginnings. She
had a ferocious pride of her own, but it was not of the kind that
meanly denies its origin.
“Father,” was her gay greeting to the astonished and still coatless
Joe, “I’ve brought somebody to see you.”
Jack, wearing a dustcoat and other appurtenances of the
chauffeur’s craft, had followed upon the heels of Princess Bedalia
into the front parlor of Number Five. In response to the young man’s
bow, Kelly offered a rather dubious hand. As became a symbol of
law and order and a member of the straitest sect of the Pharisees,
he didn’t feel inclined to encourage Mary in gallivanting up and down
the land. Nor did he feel inclined to give countenance to any
promiscuous young man she might bring to the house.
“Mr. Dinneford—my father, Police-Sergeant Kelly.” It was a
delightfully formal introduction, but rather wickedly contrived.
Jack was so taken aback that he felt as if a feather might have
downed him. But even to the lynx eyes of Mary, which were covertly
upon him, not a trace of his feelings was visible. He merely bowed a
second time, perhaps a little more gravely than the first.
“Pleased to meet you, sir,” said Sergeant Kelly, in a voice which
showed pretty clearly that he was overstating the truth.
Mary could not repress the rogue’s laugh that sprang to her lips.
“Where’s my old mumsie?” she gayly demanded, partly in the
hope of concealing her wicked merriment.
“Upstairs with your Aunty Harriet.”
“Aunt Harriet here!” The tone was full of surprise. And then the
charming voice took a turn affectionately non-committal. “What luck!
It seems an age since I saw her.”
In spite of himself, Joe could not help being a little in awe of the
girl. She was so remarkably striking that every time he saw her it
became harder to keep up the pretense of blood relationship. She
had developed into the finest young woman he had ever met. Her
official father was very proud of her, the affection she inspired in him
was true and real, but at the moment he was more than a little
embarrassed by the impact of an immensely distinguished
personality.
However, in spite of such beauty and charm, he was determined
to do his duty by her; as became a father and a man he felt bound to
admonish her.
“Since you took up with those people, none of us have been
seeing much of you,” he forced himself to say, in his most magisterial
manner.
“Old story!”
“It’s true and you know it.” Joe declined on principle to be
softened by her blandishments.
“Wicked old story!” She took him by the shoulders and shook
him; and then she sighed as a mother might have done, and gazed
into his solemn face. “Father,” she said, “you are an old and great
dear.”
“Get along with you!” said Joe sternly, but in spite of himself he
couldn’t help laughing.
“I’ll leave you and Mr. Dinneford to have a little crack while I take
this to my mumsie.” Brandishing an important-looking milliner’s box,
she left the room in a laughing search of Eliza.
As soon as Jack found himself alone with Mary’s father a period
of constraint ensued. It would have been wrong to deny that his
reception had been the reverse of cordial. The sensitiveness of a
lover, in duty bound to walk delicately, made no secret of that.
Moreover, he was still so astonished at Mary’s paternity that he felt
quite at a loss. Nature had played an amazing trick. Somehow this
serio-comic London copper in half-mufti, was going to make it very
difficult to exercise the deference due to a prospective father-in-law.
An acute silence was terminated by Joe’s “Won’t you sit down,
sir?”
Jack sat down; and then Mary’s father, torn between stern
disapproval and the humane feelings of a host, invited the young
man solemnly to a glass of beer.
“Thank you very much,” said Jack, with admirable gravity.
Murmuring “excuse me a minute,” Joe went to draw the beer. Left
alone the young man tried to arrange his thoughts; also he took
further stock of his surroundings. He had yet to overcome a powerful
feeling of surprise. It was hard to believe that Princess Bedalia, in
the view of her fiancé, the very last word in modern young women,
should have sprung from such a milieu as Number Five,
Beaconsfield Villas. It was a facer. Yet somehow the chasm between
Mary and her male parent seemed almost to enhance her value. She
was so superb an original that she defied the laws of nature.
The young man was engulfed in an odd train of speculation when
Mary’s father returned with the beer. He poured out two glasses,
gave one to the visitor, took one himself, and after a solemn “Good
health, sir!” solemnly drank it.
Jack returned the “Good health!” and followed the rest of the
ritual. And then feeling rather more his own man, he made an effort
to come to business. But it was only possible to do that by means of
a directness verging upon the indelicate.
“Sergeant Kelly,” he said, “have you any objection to my marrying
Mary?”
No doubt the form of the question was a little unwise. At least it
exposed the young man to the prompt rejoinder:
“I know nothing whatever about you, sir.”
“My name is Dinneford”—he could not refrain from laughing a
little at the portentous gravity of a prospective father-in-law. “And I
think I can claim that I have always passed as respectable.”
“Glad to hear it, sir,” said Joe, the light of a respectful humor
breaking upon him. And then measuring the young man with the eye
of professional experience. “May I ask your occupation?”
“No occupation.”
“I don’t like the sound o’ that.” Sergeant Kelly sagely shook his
head.
“Perhaps it isn’t quite so bad as it sounds,” said the young man.
“At present, you see, I am a kind of understudy to a sort of uncle I
have. I am in training as you might say, so that one day I may follow
in his footsteps.”
“An actor,” said the dubious Joe. He didn’t mind actors
personally, but impersonally he didn’t quite hold with the stage.
“Not exactly,” said the young man coolly, but with a smile. “And
yet he is in his way. In fact, you might call him a prince of
comedians.”
“I’m sorry, sir.” Sergeant Kelly measured each word carefully. “But
I’m afraid that’s only a very little in his favor.”
“I’m sorry, too,” said Jack. “My uncle is a duke, and the deuce of
it is, I have to succeed him.”
“A duke!” Sergeant Kelly’s tone of rather pained surprise made it
clear that such a romantic circumstance greatly altered the aspect of
the case. It also implied that he was far from approving an ill-timed
jest on a sacred subject. His brow knitted to a heavy frown. “Well, sir,
I can only say that if such is the case you have no right to come a-
courting our Mary.”
“For why not, Sergeant Kelly?”
“You know why not, sir, as well as I do. She’s a fine gal, although
I say it who ought not, but that will not put her right with your friends.
They will expect you to take a wife of your own sort.”
“But that’s rather my look-out, isn’t it?”
“Yes, sir, it is,” said Joe, with the air of a warrior, “but as you have
asked me, there’s my opinion. The aristocracy’s the aristocracy, the
middle-class is the middle-class, and the lower orders are the lower
orders—there they are and you can’t alter ’em. At least, that’s my
view of the matter.”
Jack forced a wry smile. Mary was a chip of the old block. Such
an uncompromising statement seemed at any rate to explain the
force of her conviction upon this vexed subject.
“Excuse the freedom, sir,” said the solemn Joe, “but you young
nobs who keep on marrying out of your class are undermining the
British Constitution. What’s to become of law and order if you go on
mixing things up in the way you are doing?”
The young man proceeded to do battle with the Philistine. But the
weapons in his armory were none of the brightest with which to meet
the crushing onset of the foe.
“It’s no use, sir. As I say, the aristocracy’s the aristocracy, the
middle-class is the middle-class, and the lower orders are the lower
orders—there they are and you can’t alter ’em. You don’t suppose
I’ve reggerlated the traffic at Hyde Park Corner all these years not to
know that.”
In the presence of such a conviction, the best of Jack’s
arguments seemed vain, futile and shallow. Fate had charged
Joseph Kelly with the solemn duty of maintaining the fabric of
society, and in his purview, no argument however cunning, could set
that fact aside.
IV
While these two were still at grips, each meeting the arguments
of the other with a sense of growing impatience, the cause of the
trouble intervened. Mary came into the room, leading her mother by
the hand. With the face of a sphinx followed Harriet.
The blushing Eliza was adorned with a fine coat which had come
in the milliner’s box. Mary had laughingly insisted on her mother
appearing in it, in spite of Eliza’s firm conviction that “it was much too
grand.”
“My word, mother!” roared Joe, at the sight of her splendor. “I’m
thinking I’ll have to keep an eye on you.”

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Biological fluid surface interactions in detection

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The quantitative detection of species of biological interest in blood, serum,

RSC Detection Science Series No. 8

Finally, we especially would like to express our deep appreciation to

Michael Thompson, Sonia Sheikh,

The Physical Chemistry of Blood–Surface Interactions

RSC Detection Science Series No. 8

surfaces; this is the primary focus of this book.

Examples of approaches are then presented using surface chemistry, morph-

nostics (or in their combined form as ‘theranostics’). It also addresses con-

Chapter 1 Relevant Aspects of Surface Physical Chemistry 1

Chapter 2 Protein Adsorption on Surfaces: Understanding the

RSC Detection Science Series No. 8

2.2 Structural Complexity and Diversity of Proteins 48

Chapter 3 Interaction of Cells and Tissue with Substrate Surfaces 81

Chapter 4 Biological Consequences of the Blood–Surface

4.1.2Blood Coagulation and Blood–Material

4.1.4 Complement System 152

Chapter 5 Antifouling Surface Chemistries to Minimize Signal

Chapter 6 Prevention of Deleterious Biofluid–Surface Interactions

6.1.2 Chemical View: the Pivotal Cloaking Role of

through both Empirical and Computational

Subject Index 281

Relevant Aspects of Surface

RSC Detection Science Series No. 8

physical sensors and the implementation of novel sensing principles, and on

1.1.1 Materials and Biological Systems – The Biomaterials

Relevant Aspects of Surface Physical Chemistry 3

surface. This non-specific adsorption, called fouling, can dominate the

1.1.2 The Problem of Fouling

1.2 Interfaces and Their Properties

Precise definitions of the main interfacial properties are imperative since

1.2.1 Definition and Thermodynamic Aspects

Relevant Aspects of Surface Physical Chemistry 5

represents the internal energy of the interface region. Analogous equations

U a/b ¼ V a/bu a/b (1.3)

However, as mentioned previously, the change in a property that takes place

Relevant Aspects of Surface Physical Chemistry 7

according to the Gibbs criterion. It is useful to define the intensive interface

1.2.2 Surface Tension and Surface Free Energy

The proportionality factor g is known as the surface tension of the liquid,

Relevant Aspects of Surface Physical Chemistry 9