HPCT311:LABORATORY
WEEK 1: LABORATORY SAFETYAND KINDS OF MICROSCOPE
2nd Semester I S.Y. 2022-2023
TRANSCRIBED BY: TUGUINAYO
LABORATORY SAFETY, INSTRUMENTATION AND QUALITY MANAGEMENT
SYSTEM IN HISTOPATHOLOGIC LABORATORY
Histopathology
• art of analyzing and interpreting the shapes, sizes and architectural
patterns of cells and tissues within a given specific clinical background
• Activities: • pre-analytical • analytical • post-analytical
RISK MANAGEMENT AND SAFETY IN THE LABORATORY
• Risk management
• process of ensuring and maintaining personal as well as environmental
health and safety in the laboratory
• The first step is to identify all electrical, mechanical and biological hazards
that can potentially cause harm in the laboratory.
• An inventory of chemical reagents must be on hand and obsolete chemicals
should be routinely disposed of.
• The following are general safety precautions that must be observed when
working in the laboratory:
• Protect the hands and forearms by wearing either gloves and a laboratory
coat or suitable long gloves to avoid contact of the toxic material with the
skin. Wash hands frequently throughout the day and before leaving the lab.
• Procedures involving volatile toxic substances and those involving solid or
liquid toxic substances that may result in the generation of aerosols should
be conducted in a fume hood or other suitable containment device.
• The laboratory workplace should be well-ventilated, clean and organized.
Smoking, sleeping, eating and drinking are prohibited in the laboratory. Do
not store food and drinks in laboratory refrigerators. Do not wear shorts,
sandals, or open-toed shoes in laboratory.
• Minors or personal pets are not permitted in the laboratory.
• Secure any dangling jewelry, restrain loose clothing, and tie back long hair
that might get caught in equipment before starting work.
• Use of cell phones and music headphones should be avoided while working
in the lab. They can be distracting and can increase the potential for an
accident to occur. They can also become contaminated if handled while
working with hazardous materials.
• Every instrument used in the laboratory should meet electrical safety
specifications and have written instructions regarding its use.
• Eye wash station, safety shower and first aid kits should be standard
facilities in a laboratory. Fire extinguishers, emergency shower systems,
emergency eye washers, first aid, emergency blankets, and hoods must be
checked monthly.
• To avoid the unnecessary purchase of chemical materials, a detailed list of
chemical materials must be prepared. Only a minimum amount of volatile
chemicals must be kept in the laboratory.
• Chemical material should be stored and safely secured where there is
sufficient ventilation. Combustible chemical material must be stored in a heat
resistant cabinet. Acids and bases must be separately stored.
• Every chemical compound used in the laboratory should have a materials
safety data sheet on file that specifies the nature, toxicity, and safety
precautions to be taken when handling the compound.
• All chemical material must be labeled with the name, characteristics,
danger level, and precautionary measures.
• Laboratories must have available appropriate protective gears for all
individuals: safety devices, goggles, gloves, lab coats, and face-shields.
• The laboratory must have a method for disposal of hazardous wastes.
Collect and seal absorbed material into labelled containers for disposal.
Tissues that are collected should be stored in formalin and may be disposed
by incineration or by putting them through a "tissue grinder" attached to a
large sink (similar to a large garbage disposal unit). Used chemicals must not
be released into soil, drains and waterways. Use an absorbent such as sand,
“kitty litter” or a commercial product to collect spills and contain spread.
• One must always be cautious when handling electrical appliances and must
be aware of the location of safety devices (fire extinguisher, emergency
shower system). Extinguishers with water, carbon dioxide, dry chemical
powder or foam are all suitable depending on other products involved in a
fire. Fire safety procedures should be posted. There must not be any obstacle
in the vicinity of the laboratory door.
• Avoid handling the sharp ends of instruments. Use forceps or other tools to
remove sharp instruments from baskets and autoclaves. Workers should use
appropriate hand protection when hands are exposed to hazards such as
cuts, lacerations or thermal burns.
• Laboratory accidents must be documented and investigated with incident
reports and industrial accident reports. Obtain medical advice (first aid
officer, doctor, poisons information center, ambulance) immediately if major
exposure occurs.
TYPES OF HAZARDS
• Chemical Hazards
• Physical Hazards
• Biological Hazards
Chemical Hazards
• Cleaning agents and disinfectants, drugs, anesthetic gases, solvents, paints,
and compressed gases
• Potential exposures to chemical hazards can occur both during use and
with poor storage
• The “lab standard” applies to the laboratory use of chemicals and
mandates written in the Standard Operating Procedures (SOPs) that address
the particular hazards and precautions required for safe use.
• Permissible Exposure Limits (PELs), Threshold Limit Values (TLVs), or
Occupational Exposure Limits (OELs) are some of the terms used to define
the maximum allowable airborne concentration of a chemical (vapor, fume
or dust) to which a worker may be exposed
➢ Labeling
• Every chemical should be labeled with certain basic
information, including: Chemical name and, if a mixture,
names of all ingredients;
• Manufacturer's name and address if purchased
commercially, or name of person making the reagent;
• Date purchased or made;
• Expiration date, if known;
• Hazard warnings and safety procedures.
The different types of chemicals include:
1. Irritants are chemicals that cause reversible inflammatory effects at the
site of contact with living tissue, especially the skin, eyes and respiratory
passages.
2. Corrosive chemicals cause destruction or irreversible alterations when
exposed to living tissue, or destroy certain inanimate surfaces (generally
metal). A chemical may be corrosive to tissue but not to steel, or vice- versa.
Few are corrosive to both.
3. Sensitizers cause allergic reactions in some exposed workers, not just in
hypersensitive individuals. Sensitization may occur at work because of the
high exposure level.
4. Carcinogens are substances that induce tumors, not only in experimental
animals but also in humans. Examples of carcinogenic chemicals include
chloroform, chromic acid, formaldehyde, nickel chloride and potassium
dichromate. Carcinogenic dyes include auramine, basic fuchsin, and any dye
derived from benzidine (including Congo red and diamino- benzidine). 5.
Toxic materials are capable of causing death by ingestion, skin contact or
inhalation at certain specified concentrations. These include methanol,
chromic acid, osmium tetroxide and uranyl nitrate.
Physical Hazards
• slips and falls from working in wet locations and the ergonomic hazards of
lifting, pushing, pulling, and repetitive tasks. Other physical hazards often
unnoticed are electrical, mechanical, acoustic, or thermal in nature. Ignoring
these can have potentially serious consequences.
• Many operations in the lab can result in lab workers assuming sustained or
repetitive awkward postures such as looking at slides on a microscope for
extended periods
HPCT311:LAB 1
Biological Hazards
• refer to anything that can cause disease in humans, regardless of their
source.
• Biohazards include infectious agents and their toxins as well as
contaminated solutions, specimens or objects.
• Allergens, are one of the most important health hazards, yet they are
frequently overlooked.
• Molds and fungi produce and release millions of spores small enough to be
air, water, or insect- borne which may have negative effects on human health
including allergic reactions, asthma, and other respiratory problems.
USE AND CARE OF THE MICROSCOPE
• microscope is one piece of equipment that is used by both the pathologist
and the histotechnologist.
• pathologist examines the slide under the microscope to identify a disease
process or an abnormality that will directly affect the patient's treatment.
•histotechnologist examines the same slide microscopically for quality
control to determine whether all technical processes are done properly and if
a slide of diagnostic quality has been achieved.
microscope must accomplish three things:
(1) it must magnify the object
(2) it must resolve the details of the object
(3) it must make these details visible.
COMPOUND MICROSCOPE
• microscope with more than one lens and its own light source. In this type of
microscope, there are ocular lenses in the binocular eyepieces and objective
lenses in a rotating nosepiece closer to the specimen. Because it contains its
own light source at its base, a compound light microscope is also considered
a bright field microscope, which means that the specimen is lit from below
and viewed from above. Illumination comes from below and contrast in the
sample is caused by absorbance of some of the transmitted light in dense
areas of the sample.
➢ Viewing Heads
• Monocular Heads - only use one eyepiece when viewing the
specimen.
• You are restricted if you want to use an LCD camera because this
would occupy the eyepiece. However, monocular microscopes are
light weight and are inexpensive.
• Binocular heads have two eyepieces and are more convenient
and comfortable to use. It is the most common choice.
• Trinocular Heads - have a third eyepiece tube that can be used
by another person simultaneously or by an LCD camera. The
trinocular option is more expensive than the other two types.
The main framework of the Compound Microscope consists of:
• Base -provides support for the microscope. The base should be large and
solid enough to allow the microscope to stand by itself.
• Arm - supports and holds the magnifying and adjustment system. It can be
used as a handle for carrying the microscope.
• Stage - is the flat platform where the slide is placed for examination.
• Substage - is located directly under the stage and holds the condenser
• and diaphragm.
• Mechanical Stage - permits movement of the stage while holding the
slide in the phase of focus.
The parts of the lens system are:
1. Nosepiece - is located at the end of the body tube for holding the
objectives.
2. Objectives - consist of a system of lenses located at the end of the body
tube that is held in place by the nosepiece and is closer to the slide under
examination. The purpose of the objective is to increase or decrease
magnification. The objectives are mounted on a revolving turret allowing for
the change of objectives. When one objective is focused on the turret, all
lenses will be approximately in focus. If this is true, the microscope is said to
be Par focal.
3. Focal length - is the distance between outer lens of objective and the
cover glass of the slide under examination.
COMPOUND MICROSCOPE
• Magnification is the process that increases the size of the
structure under examination. It is achieved by the use of the
microscope's lens system. The total magnification of a microscope
is the product of the magnifying power of the objective and
eyepiece, with a normal tube length of 160 mm.
➢ Bright field microscopy
• generally used in compound microscopes, where light is either
passed through, or reflected off, a specimen.
• Illumination is not altered by devices that alter the properties of
light
• "bright field" is derived from the fact that the specimen is dark
and contrasted by the surrounding bright viewing field.
• Simple light microscopes are sometimes referred to as bright
field microscopes where a specimen is placed on the stage of the
microscope and incandescent light from the microscope’s light
source is aimed at a lens beneath the specimen.
• This lens is called a condenser.
• The condenser usually contains an aperture diaphragm to
control and focus light on the specimen; light passes through the
specimen and then is collected by an objective lens situated in a
turret above the stage.
• The objective magnifies the light and transmits it to an oracular
lens or eyepiece and into the user’s eyes
➢ Dark Field illumination
• technique used to observe unstained and transparent samples
causing them to be clearly visible and appear brightly lit against a
dark, almost purely black background.
• A dark field microscope blocks this central light with a
condenser so that only oblique rays hit the object. The principal
elements of dark field illumination are the same for both
stereomicroscopes and more conventional compound
microscopes. Illumination of specimens by dark field requires
blocking out of the central light rays along the optical axis of the
microscope, which ordinarily pass through and around
(surrounding) the specimen. Blocking these light rays allows only
those oblique rays originating at large angles to strike the
specimen positioned on the microscope stage.
➢ Phase contrast microscopy
• optical microscopy illumination technique in which small phase
shifts in the light passing through a transparent specimen are
converted into amplitude or contrast changes in the image.
• type of light microscopy that enhances contrasts of transparent
and colorless objects by influencing the optical path of light
• phase contrast microscope is able to show components in a cell
or bacteria which would be very difficult to see in an ordinary light
or bright field microscope.
➢ Polarized light microscopy
• contrast-enhancing technique that improves the quality of the
image obtained with birefringent materials when compared to
other techniques such as bright field microscopy, phase contrast
microscopy and dark field microscopy.
• It is designed to examine specimens that are visible primarily
due to their optically anisotropic character.
• two essential components:
(1) The polarizer situated below the specimen stage usually fixed
in the left-to-right, East-West direction, although this is usually
rotatable through 360 degrees.
(2) The analyzer, usually aligned North-South but again rotatable
on some microscopes, is located above the objectives and can be
moved in and out of the light path as required.
HPCT311:LAB 2
 • participating laboratories are asked to stain sections that have
FLUORESCENCE MICROSCOPE been submitted by the scheme organizer
• refers to any microscope that uses fluorescence to generate an Continuing quality improvement
image. • system is used to approach, evaluate and identify opportunities to improve
• In fluorescence microscopy, many wavelengths of light, ranging quality before problems occur through evaluation of all systems/processes in
from the ultraviolet to the visible can be used to cause samples to the laboratory
• goal: improve potential care and safety through recognition of potential
fluoresce and allow viewing by eye or with the use of specifically
problems/errors before they can occur
sensitive cameras. • When certain compounds are illuminated
with high energy light, they emit light of a lower frequency. Pre-analytical phase
• This effect is known as fluorescence. • sample collection, transport, accession and tissue processing and
submission of the slide for reporting
ELECTRON MICROSCOPE • can endanger quality of histopathology report
• microscope that uses a beam of accelerated electrons as a
source of illumination. Analytical phase
• Because the wavelength of an electron can be up to 100,000 • component:
times shorter than that of visible light photons, the electron • slide reading along with relevant data and preparation of report
microscope has a higher resolving power than a light microscope
and can reveal the structure of smaller objects. Post-analytical aspects
• follow the analytical phase
➢ Transmission Electron Microscope (TEM)
• components:
• works on the same principle as an optical microscope but uses
• preparation and delivery of the report
electrons instead of light and electromagnets instead of glass • archiving of request form and report
lenses. • storage of the reported specimen for set retention period and safe disposal
• Use of electrons instead of light allows a much higher resolution, of specimen
giving a two- dimensional view.
• Thin slices of specimen are obtained.
• The electron beams pass through this. It has high magnification
and high resolution.
• A transmission electron microscope can achieve better than 50
nm resolution and magnifications of up to about 10,000,000 x
whereas most light microscopes are limited by diffraction to
about 200 nm resolution and useful magnifications below 2000x.
➢ Scanning Electron Microscope
• uses electron illumination and looks at the surface of bulk
objects by scanning the surface with a fine electron beam.
• image is seen in 3-D.
• It has high magnification and high resolution. • The specimen is
coated in gold and the electrons bounce off to give you an
exterior view of the specimen.
• The pictures are in black and white.

Laboratory Quality Management System

Quality
• degree to which healthcare services strive to provide accurate desired
outcomes for patients and are consistent with current professional
knowledge
Safety
• “freedom from accidental injury”
• hazards: toxic chemicals, pathogenic microorganisms, mechanical, electrical
and fire
• safety consciousness & safety practices
Quality control
• system of routine technical activities
• provides routine and consistent checks to identify, address errors and
omissions, ensures data integrity, correctness and completeness and also
records all quality control activities
Quality Assurance
• planned system of review procedures conducted by personnel not directly
involved in the laboratory process
• data of QC provides the data for QA
• correlation of errors, complaints, failures or other unexpected results
• Quality assessment programs
• College of American Pathologists (CAP)
• United Kingdom National External Quality Assessment Service (UK NEQAS)
➢ two distinct systems:
• selective system:
• stained preparations from departmental archival records are
used to assess the quality of staining
• distributive system

HPCT311:LAB 3