RISK MANAGEMENT AND SAFETY IN THE

LABORATORY
Saturday, October 8, 2022

RISK MANAGEMENT - Pertains to the 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
→ Standard operating procedures must be detailed to include control of hazardous
substances, risk assessments, and other health and safety information relevant to
handling of specimens
→ One of the most common accidents in the laboratory involves cutting one's finger or hand
on microtome knives
→ The risk manager should develop a system whereby all incidents and accidents are
reported, no matter how small. Each incident should be investigated and, where possible,
additional measures should be taken to ensure that the incident does not happen again

GENERAL SAFETY PRECAUTIONS

→ Many laboratories contain significant risks, and the prevention of laboratory accidents
requires great care and constant vigilance

→ The laboratory environment can be a hazardous place to work. Laboratory workers are
exposed to numerous potential hazards including chemical, biological, physical, and
radioactive hazards, as well as musculoskeletal stresses

→ All laboratories need to have a written program stating the policies, procedures, and
responsibilities that serve to protect employees from the health hazards associated with
that particular workplace

→ Measures to protect against laboratory accidents include safety training and

enforcement of laboratory safety policies

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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, 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 cellphones and music headphones should be avoided while working in the lab
• 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.

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 • 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

OCCUPATIONAL HAZARDS
Health Hazard - The potential to cause harem to health
▪ Chemical
▪ Physical
▪ Biological
▪ Ergonomic

OCCUPATIONAL HEALTH HAZARDS

Physical Biological

- Noise - Bacteria
- Vibration - Fungi
- Radiation - Insects
- Lightning - Viruses
- Heat
- Cold
- Barometric pressure
Chemical Ergonomic

- Fumes - Manual handling

- Gases - Repetitive movement
- Aerosols - Restrictive working area
- Mists - Prolonged exertion
- Liquids - Job stress
- Dusts - Job monotony
- Unsociable hours
- Job organization
- Violence / harassment

SAFETY HAZARDS
• Anything that can have an adverse impact on one's safety
• Associated with instantaneous events that have adverse effects on the work
process accidents

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 • May involve a health hazard but is concerned with how their effects are realized due to:
→ Non-ideal workplace conditions
→ Risky acts (not following protocols / process)

CHEMICAL HAZARD
• Cleaning agents and disinfectants, drugs, anesthetic gases, solvents, paints, and
compressed gases are examples of chemical hazards.
• The potential for harm or injury could be significant if chemicals are misused or
mishandled. 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.
• Explosive chemicals include picric acid. Certain silver solutions may explode upon aging,
which is why they should never be stored after use. Oxidizers are harmless by
themselves, but may initiate or promote combustion and present a serious fire risk when
in contact with certain substances. Examples include sodium iodate, mercuric oxide and
chromic acid.
• 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. While they represent a concentration at or below which it is safe for most
people to work, some individuals may react adversely even below such limits due to
hypersensitivity or allergy.

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.

DIFFERENT TYPES OF CHEMICALS

a. Irritants are chemicals that cause reversible inflammatory effects at the site of contact
with living tissue, especially the skin, eyes and respiratory passages.
b. 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.
c. Sensitizers cause allergic reactions in some exposed workers, not just in hypersensitive
individuals. Sensitization may occur at work because of the high exposure level.
d. 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).
e. 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.

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STORAGE OF HAZARDOUS CHEMICALS
• Standard precautions will provide laboratory workers with good protection from most
toxic substances. In addition, records that include amounts of material used and names
of workers involved should be kept as part of the laboratory notebook record of the
project.
• To minimize hazards from accidental breakage of apparatus or spills of toxic substances
in the fume hood, they should be stored in pans or trays made of polyethylene or other
chemically resistant material. The apparatus should be mounted above trays of the same
type of material.

PHYSICAL HAZARDS
• The most obvious physical hazards are 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. What
is found acceptable for brief or occasional use may become problematic if performed for
long durations or very frequently. Pain is a good indicator that something is wrong. Work
must be conducted in a neutral, balanced posture.

ELECTRICAL HAZARDS
• In the laboratory, workers may be exposed to electrical hazards including electric shock,
arc blasts, electrocution, fires and explosions. Potential exposures to electrical hazards
can result from faulty electrical equipment/instrumentation or wiring, damaged
receptacles and connectors, or unsafe work practices. Damaged electrical cords can
lead to possible shocks or electrocutions.

BIOLOGICAL HAZARDS
• Biohazards 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.
• If biological materials are used in the area, they should not be stored in hallways, in
unlocked freezers or in refrigerators. Biohazard signs should be placed in appropriate
areas.
• Pathologists, histotechnologists and technicians may be exposed to a certain level of risk
when handling and processing potentially infectious specimen through inhalation of
aerosols, contact with non-intact skin and contact with mucous membranes (eyes, nose,
mouth). Fresh tissue and body fluids must always be considered potentially infectious,
and grossing of specimen has the highest risk of all histological activities. Fixed
specimens have a much less risk because nearly all infectious agents are deactivated by
histological fixation, although tissues must be thoroughly fixed for this to happen.
Complete penetration by alcohol will destroy all infectious agents except prions.

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 • Prions are infectious agents that cause spongiform encephalopathies such as
Creutzfeld-Jakob disease (CJD), scrapie and mad cow disease. Normal steam
sterilization does not inactivate these particles, and common effective treatments
• like sodium hypochlorite or phenol will create artefacts in tissue. Tissue from patients
with suspected CJD can be decontaminated by immersing the specimen in formalin for
48 hours, followed by treatment in concentrated formic acid for 1 hour, and additional
formalin fixation for another 48 hours.
• Small dust-like particles generated from sectioning may become airborne, particularly
when performing cryostat sections of fresh tissue. Cryogenic sprays can magnify this
risk, and therefore should not be used to freeze potentially infectious tissue. Cutting
areas or surfaces may be sterilized with chlorine bleach or a suitable commercial
disinfectant, and warning signs should be posted in labs handling infectious materials.
Disinfectants should be on hand for sanitizing bench tops and treating spills. Biological
safety cabinet(s) must be certified within the last 12 months of use.

HANDLING SPILLS
• The laboratory worker must be prepared for potential accidents or spills involving toxic
substances. Lab workers must be trained in handling toxic materials and spill clean-up
before beginning work with toxic substances.
• Small spills are defined as those that can be safely handled by the immediate staff. Spill
neutralizing and containment kits should be available immediately outside the hazardous
work area. These may be commercially purchased or assembled from common
materials, and should include protective equipment and clean up aids, such as good
quality latex or nitrile gloves similar in thickness to dishwashing gloves, disposable
plastic aprons for chemical spills and disposable gowns for biohazards, dustpan and
brush for powders, sponges, towels and mops for liquids, adsorbent material (kitty litter
or a commercial sorbent), bleach (sodium hypochlorite for biohazards), baking soda for
acids, vinegar (5% acetic acid) for alkalis, a commercial neutralizing product, a sealable
plastic bucket and heavy plastic bags for containment of the salvaged waste.
• If the amount of spilled material is limited to a few grams or milliliters, it can be simply
wiped off with towel or sponge, while protecting the hands with suitable gloves. The
towel or sponge must be disposed of appropriately after use; do not put it into the
general trash, and protect the room from its vapors by sealing it within an impermeable
plastic bag or container.
• For significant spills of dangerous materials, all personnel should evacuate the room or
immediate vicinity where the accidental spilled occurred, and first aid must be given to
anyone who has gotten splashed or is feeling the effects of vapors. If the spill is large,
the area must be sealed off and an experienced emergency response team must be
called.
• If there is a major spill outside of the hood, the room or appropriate area
• should be evacuated and necessary measures should be taken to prevent exposure of
other workers. Spills must be cleaned by personnel wearing suitable personal protective
apparel. If a spill of a toxic material occurs outside the hood, an air-supplied full-face
respirator may be needed. The work space and equipment should be decontaminated
with 10% bleach solution. Avoid creating dust. Contaminated pipette tips, tubes, weighing
trays, gloves, paper towel, napkins and any other clean up debris must be disposed of as
hazardous waste. After removal of gloves, wash hands thoroughly with soap and copious
amounts of water.

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FIRST AID MEASURES
With laboratory chemicals, the most common accidents requiring first aid are ingestion, eye
contact and extensive skin contact.

Laboratory technicians and technologists should have basic training in dealing with these
situations, and yearly safety training should include first aid information and preparedness in
the event of chemical accidents, including accidental ingestion of hazardous chemicals.

First aid kits must be easily accessible and refilled on a regular basis.

When providing first aid treatment to a person that has been exposed to chemical, infectious
or toxic waste products, the following precautions should be observed:
→ Immediately remove the person from the source of contamination and move to fresh air.
→ If the person is not breathing, do not use mouth to mouth, or mouth to nose ventilation,
because of the danger to the rescuer. Instead, use a resuscitation bag and mask.
→ If pulse is absent, start external cardiac massage and follow standard Advanced
Cardiovascular Life Support (ACLS) guidelines.
→ Give 100% oxygen by mask if available.
→ Remove all contaminated clothing and footwear into a sealable collection bag and
launder contaminated clothing thoroughly.

For accidental skin contact with hazardous chemicals, the affected area should be washed
with copious amounts of water for 15-30 minutes. Emergency showers should be as
accessible as eye wash stations. If the hazardous substance is not readily water-soluble, use
soap with the water wash. Immediately remove contaminated clothing and launder before re-
use. Seek medical assistance following skin contact.

Splashing of dangerous chemicals into the eyes is also a common accident. Symptoms include
redness, pain, blurred vision, and eye damage. All laboratories should be equipped with
emergency eyewash stations, either as standing devices or small appliances affixed to sink
faucets. Current recommendations are to have such devices no more than 100 feet from
hazardous work areas, and the water temperature should be controlled to a range of 15-35°C.
In case of accidental splashing, the affected eye should be rinsed for 15-30 minutes, pulling
the lids away from the eyeball, prior to seeking emergency health care. Portable eyewash
bottles are not recommended because they pour too little liquid and may become
contaminated with microorganisms.

ERGONOMICS
Laboratory work activities can introduce ergonomic risk factors that are associated with
muscular-skeletal disorders. Laboratory-associated ergonomic risks are the same as those
found in the office and general industry.

These risk factors include awkward or sustained postures, highly repetitive movements,
excessive force or strain, contact stresses, and vibration.
• Awkward postures occur when body parts are positioned away from their neutral
position. These postures can put stress on the joint and its associated muscles.
• Contact stress is a sustained contact between a body part and an external object.
Examples include: resting the wrist or forearm against a sharp edge/corner.
• Duration is the period of time that a body part is exposed to an ergonomic risk factor.
Longer durations of exposure increase the severity of the risk.

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 • Repetition is the repeated performance of motion that includes other ergonomic risk
factors such as force and/or awkward posture. Severity of the risk increases with higher
repetition of motions with ergonomic risk factors.
• Static postures occur when a body part is held in a single position over a long period of
time. The severity of a static posture can increase if the posture is awkward, applies
continual force, and/or is held for long durations. Examples include: sitting or standing in
single position for a long duration.

WHEN USING A MICROSCOPE

• Sit close to the work surface.
• Avoid leaning on hard edges.
• Keep elbows close to their sides.
• Adjust chair, workbench, or microscope as needed to maintain an upright head position.
• Elevate, tilt or move the microscope close to the edge of the counter to avoid bending the
neck.
• Take short breaks. Every 15 minutes, close the eyes or focus on something in the
distance.
• Every 30-60 minutes, get up to stretch and move. Alternate between sitting and standing
positions.
• When possible, plan work tasks to include a variety of movements to avoid static
postures or repetitive motions.
• Avoid contacting or resting wrists or forearms on sharp edges.
• When seated, the thighs should be parallel to the floor and feet firmly planted on the
floor or on a foot rest.

MAINTAINING PROPER POSTURE:

• Keep your back straight, maintain all 3 natural curves in your spine.
• Distribute your weight evenly on both hips.
• Keep your head and neck aligned over your shoulders.
• Sit back in your chair; your back should be supported by the seat back.
• Adjust your chair height so that your hips are slightly higher than your knees.
• Be sure your feet are supported by the floor or a footrest.
• Avoid sitting for long periods of time; get up from your chair at least once every hour.
• Do not twist or bend your back from a seated position.

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INTRODUCTION TO HISTOPATHOLOGY
Saturday, October 8, 2022

HISTOPATHOLOGY AND CYTOPATHOLOGY

• Histopathology refers to the microscopic examination of tissue in order to study manifestations
of the disease. Specifically, in clinical medicine, histopathology refers to the examination of a
biopsy, or surgical specimen by a pathologist after the specimen has been processed and
histological sections have been placed in a glass slide.
• In contrast, cytopathology examine free cells or tissue fragments.

PURPOSE OF HISTOPATHOLOGY
1. To differentiate normal and abnormal tissues.
2. Some diseases can only be diagnosed by histopathological techniques (e.g. Bovine spongiform
encephalopathy)

DEFINITION OF TERMS
• Histology –the study of the microscopic structure of tissues.
• Histopathology –the study of changes in tissues caused by disease.
• Cytology –the branch of biology concerned with the structure and function of plant and animal
cells.
• Cytopathology – a branch of pathology that deals with manifestations of disease at the cellular
level.
• Autolysis – the destruction of cells or tissues by their own enzymes, especially those released by
lysosomes.
• Pathology - the science of the causes and effects of diseases, especially the branch of medicine
that deals with the laboratory examination of samples of body tissue for diagnostic or forensic
purposes.
• Etiology - cause or origin of disease or abnormal condition.
• Homeostasis - A property of cells, tissues, and organisms that allows the maintenance and
regulation of the stability and constancy needed to function properly.

CELLULAR ADAPTATION, INJURY, AND DEATH

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HYPERPLASIA
• response to increased demand and external stimulation
• ↑ number cells - ↑ volume of organ
• often occurs with hypertrophy

HYPERTROPHY
• response to ↑ functional demand or hormonal stimulation
• ↑ size of cells → ↑ size of organ (no new cells), due to ↑ synthesis structural components
• occurs in nondividing cells (e.g. myocardium)
• physiologic or pathologic

ATROPHY
• shrinkage in size of cells by loss of structural components of cell
• adaptive response, may lead to cell death

METAPLASIA
• reversible change where one adult cell type is replaced by another type
• may be adaptive to withstand chronic stress such as chemical or physical irritants
• most common epithelial metaplasia: columnar to squamous resp tract due to chronic irritation
(smoking) resp tract due to Vit A deficiency salivary, pancreatic, biliary stones

CELL INJURY AND CELL DEATH

reversible injury: initially functional and morphological changes that are reversible if damaging
stimulus removed
irreversible injury and death: with continuing damage o apoptosis (not always assoc with cell injury)
and necrosis (always pathological)

CAUSES OF CELL INJURY

• hypoxia - ↓aerobic oxidative respiration (incl CO, anaemia)
cf ischaemia – loss of blood supply with ↓ O2 and metabolic substances ie glucose → more
rapid/severe cell injury)
• physical agents – mechanical trauma, burns, cold, pressure, radiation, electric shock o chemical
agents (gluc/salt if hypertonic, ↑↑ O2, poisons, pollutants, insecticides, CO, asbestos, drugs)
• infectious agents
• immunological reactions (anaphylaxis) o genetic derangements (Downs, Sickle cell, enzymes)
• nutritional imbalances (starvation, anorexia, obesity)

Acquired causes
1. Hypoxia and ischemia – due to reduced supply of blood or oxygen-carrying-capacity of blood (i.e.
anemia & carbon monoxide poisoning)
2. Chemicals and drugs – acts on the vital function of a cell such as membrane permeability,
osmotic homeostasis, or in functional capacity of the enzyme cofactor
3. Physical agents – Mechanical trauma; thermal trauma; radiation; rapid changes in atmospheric
pressure
4. Microbial agents– bacteria, viruses, fungi, protozoa, parasites.
5. Immunologic reactions – immune responses such as hypersensitivity reactions, anaphylactic
reactions; autoimmune diseases.
6. Nutritional Imbalance – nutritional deficiency (e.g.Marasmus, Kwashiorkor, Anemia) ; High fat diet
(Obesity)
7. Aging – the accumulation of injury caused by free-radicals over the year may be responsible for
cellular ageing.

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Genetic causes:
1. Developmental defect – group of abnormalities during fetal life due to errors in morphogenesis.
2. Cytogenic defect – chromosomal abnormalities (numerical & structural)
3. Single-gene defects (Mendelian Syndrome) – mutation of a single gene.
4. Disorders with multifactorial inheritation – genes in combination with lifestyle; caused by many
contributing factors (e.g. heart disease, diabetes)

APOPTOSIS
• Programmed cell death.
• If cells are no longer needed, they commit suicide by activating an intracellular death program.
This process is therefore called programmed cell death, although it is more commonly
called apoptosis (from a Greek word meaning “falling off,” as leaves from a tree).

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