A.

COMPOUND MICROSCOPE - A compound microscope works on the principle that

when a tiny object to be magnified is placed just beyond the focus of its objective lens, a virtual,
inverted and highly magnified image of the object is formed at the least distance of distinct
vision from the eye held close to the eye piece.

B. SCANNING ELECTRON MICROSCOPE - The signals used by a scanning electron

microscope to produce an image result from interactions of the electron beam with atoms at
various depths within the sample. Various types of signals are produced including secondary
electrons (SE), reflected or back-scattered electrons(BSE), characteristic X-rays and light
(cathodoluminescence) (CL), absorbed current (specimen current) and transmitted electrons.
Secondary electron detectors are standard equipment in all SEMs, but it is rare for a single
machine to have detectors for all other possible signals.

B. Transmission Electron Microscope - Electrons are made to pass through the specimen and
the image is formed on the fluorescent screen, either by using the transmitted beam or by using
the diffracted beam.
- In the transmission electron microscope, as shown in figure 1 (Nixon 1962), the electron gun at
the top illuminates the specimen with the beam angle controlled by the condenser lens. The
lenses below the specimen are used to magnify the image of the specimen which is viewed on
the final screen at many thousand times magnification. If a second electron gun is placed below
the fluorescent screen at the bottom of the column and the electron beam is projected upwards
through the same lenses this second electron source will be reduced in size by the same
amount that the specimen image is magnified. This effect can be observed with both electron
guns on at the same time and demonstrates the reversibility of rays through electron lens
systems. In this way the resolved point in the specimen image on the fluorescent screen or on
the photographic plate is equal to the focused electron probe in the plane of the specimen.
C. Scanning Electron Microscope –
D. Scanning Probe Microscope - Scanning Probe Microscopy ( SPM ) scans an atomically
sharp probe over a surface, typically at a distance of a few angstroms or nanometres. The
interaction between the sharp probe and surface provides 3D topographic image of surface at
the atomic scale. Compared with other instruments that open a window to a world of molecule-
sized spaces, SPMs are relatively simple, inexpensive, and easy to operate. Especially
appealing about the proximal probes is their multipurpose nature that offers not only a view of
individual atoms but also ways to pick them up, move them around, and position them at will.
E. Scanning Tunnelling Microscope - Scanning tunneling microscope (STM), type of microscope
whose principle of operation is based on the quantum mechanical phenomenon known as
tunneling, in which the wavelike properties of electrons permit them to “tunnel” beyond the
surface of a solid into regions of space that are forbidden to them under the rules of classical
physics. The probability of finding such tunneling electrons decreases exponentially as the
distance from the surface increases. The STM makes use of this extreme sensitivity to distance.
The sharp tip of a tungsten needle is positioned a few angstroms from the sample surface. A
small voltage is applied between the probe tip and the surface, causing electrons to tunnel
across the gap. As the probe is scanned over the surface, it registers variations in the tunneling
current, and this information can be processed to provide a topographical image of the surface.
F. Atomic Force Microscope - The Atomic Force Microscope is a kind of scanning probe microscope in
which a topographical image of the sample surface can be achieved based on the interactions between a
tip and a sample surface. The atomic force microscope was invented by Gerd Binning et al. in 1986 at
IBM Zurich based on the STM (Scanning Tunneling Microscope) already presented in 1981. While the
latter depends on the conductive samples, the AFM allows also the use of non-conductive samples. In
1987, the inventors were awarded the Nobel Prize in Physics for the achievements. A typical AFM
consists of a cantilever with a small tip (probe) at the free end, a laser, a 4-quadrant photodiode and a
scanner. The surface characteristics can be explored with very accurate resolution in a range of 100 μm
to less than 1 μm.

G. Phase Contrast Microscope - Highly refractive structures bend light to a much greater angle
than do structures of low refractive index. The same properties that cause the light to bend also
delay the passage of light by a quarter of a wavelength or so. In a light microscope in bright field
mode, light from highly refractive structures bends farther away from the center of the lens than
light from less refractive structures and arrives about a quarter of a wavelength out of phase.

Light from most objects passes through the center of the lens as well as to the periphery. Now if
the light from an object to the edges of the objective lens is retarded a half wavelength and the
light to the center is not retarded at all, then the light rays are out of phase by a half wavelength.
They cancel each other when the objective lens brings the image into focus. A reduction in
brightness of the object is observed. The degree of reduction in brightness depends on the
refractive index of the object.
- Unstained living cells absorb practically no light. Poor light absorption results in extremely
small differences in the intensity distribution in the image. This makes the cells barely, or not at
all, visible in a brightfield microscope. When light passes through cells, small phase shifts occur,
which are invisible to the human eye. In a phase contrast microscope, these phase shifts are
converted into changes in amplitude, which can be observed as differences in image contrast.
However, this label-free technique is strongly dependent on the correct alignment of
components in the optical pathway. This alignment can be disturbed by the naturally occurring
meniscus effect, causing weak phase contrast.
H. Differential Interference Microscope - DIC microscopy is a contrast-enhancing technique. DIC
uses polarized light to convert phase delays into intensity changes (contrast). The effect is
called differential, because contrast is created only in neighboring areas. Unlike in phase
contrast, the DIC image is not built globally over the entire image. Instead, adjacent structures
with different refractive indices are contrasted when in close contact with each other.
I. Fluorescence Microscope - The underlying process of fluorescence involves the absorption of
light energy (a photon) by an indicator followed by the emission of some of this light energy (as
another photon) a few nanoseconds later. Because some energy is lost in this process, the
emitted photon has less energy than the absorbed photon. Light with a short wavelength
(toward the blue) has higher energy than light with a long wavelength (toward the red).
Therefore, light emitted from an indicator usually has a longer wavelength than that of the
absorbed (excitation) light. This change is called the Stokes shift. The molecular transitions
explaining these processes can be depicted in terms of Jablonski energy diagrams.
J. Confocal Microscope - Coherent light emitted by the laser system (excitation source) passes
through a pinhole aperture that is situated in a conjugate plane (confocal) with a scanning point
on the specimen and a second pinhole aperture positioned in front of the detector (a
photomultiplier tube). As the laser is reflected by a dichromatic mirror and scanned across the
specimen in a defined focal plane, secondary fluorescence emitted from points on the specimen
(in the same focal plane) pass back through the dichromatic mirror and are focused as a
confocal point at the detector pinhole aperture.
K. Dark Field Microcope - To view a specimen in dark field, an opaque disc is placed
underneath the condenser lens, so that only light that is scattered by objects on the slide can
reach the eye (figure 2). Instead of coming up through the specimen, the light is reflected by
particles on the slide. Everything is visible regardless of color, usually bright white against a
dark background. Pigmented objects are often seen in "false colors," that is, the reflected light is
of a color different than the color of the object. Better resolution can be obtained using dark field
as opposed to bright field viewing.
L. Bright Field Microscope - With a conventional bright field microscope, light from an
incandescent source is aimed toward a lens beneath the stage called the condenser, through
the specimen, through an objective lens, and to the eye through a second magnifying lens, the
ocular or eyepiece. Most microscopes will have a built-in illuminator. The condenser is used to
focus light on the specimen through an opening in the stage. After passing through the
specimen, the light is displayed to the eye with an apparent field that is much larger than the
area illuminated. The magnification of the image is simply the objective lens magnification
(usually stamped on the lens body) times the ocular magnification.

a. http://www.funscience.in/study-
zone/Physics/OpticalInstruments/CompoundMicroscope.php#sthash.4IiZI5QV.dpbs

b. https://www.brainkart.com/article/Transmission-Electron-microscope---Principle,-
Construction,-Working,-Advantages-and-Disadvantages_6866/
The general principles of scanning electron microscopy
W. C. Nixon
Published:27 May 1971https://doi.org/10.1098/rstb.1971.0035
https://royalsocietypublishing.org/doi/abs/10.1098/rstb.1971.0035

c. https://serc.carleton.edu/research_education/geochemsheets/techniques/SEM.html
Susan Swapp, University of Wyoming

d. https://www.azonano.com/article.aspx?ArticleID=1653
Scanning Probe Microscopy ( SPM ) - Principles and Modes of Operation
Download PDF Copy
Written by AZoNanoJul 21 2006

e. Calvin F. Quate
Article Title:
Scanning tunneling microscope
Website Name:
Encyclopædia Britannica
Publisher:
Encyclopædia Britannica, inc.
Date Published:
October 28, 2013
URL:
https://www.britannica.com/technology/scanning-tunneling-microscope
Access Date: August 25, 2019

f. Principles of Atomic Force Microscopy (AFM), Arantxa Vilalta-Clementa, Katrin Gloystein,

Nikos Frangis,
Physics of Advanced Materials Winter School 2008

g. https://www.ruf.rice.edu/~bioslabs/methods/microscopy/phase.html
Phase Contrast Microscopy

https://ibidi.com/content/213-phase-contrast
Microscopy Techniques and Culture Surfaces:
Find the Perfect Match
Phase Contrast

h. https://ibidi.com/content/214-dic
Microscopy Techniques and Culture Surfaces:
Find the Perfect Match
Differential Interference Contrast (DIC)

i. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4711767/
Cold Spring Harb Protoc. 2014 Oct; 2014(10): pdb.top071795.
Published online 2014 Oct 1. doi: 10.1101/pdb.top071795
PMCID: PMC4711767
NIHMSID: NIHMS749436
PMID: 25275114
Fluorescence Microscopy
Michael J. Sanderson,1,4 Ian Smith,2 Ian Parker,2 and Martin D. Bootman3

j. https://www.olympus-lifescience.com/en/microscope-
resource/primer/techniques/confocal/confocalintro/
Thomas J. Fellers and Michael W. Davidson - National High Magnetic Field Laboratory, 1800
East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.

k. https://www.ruf.rice.edu/~bioslabs/methods/microscopy/dfield.html

l. https://www.geog.ucl.ac.uk/resources/laboratory/light-microscopy/bright-field-microscopy