HISTOPATHOLOGIC & BS MLS

3rd Year
CYTOLOGIC TECHNIQUES
LABORATORY UNIT / PROF. JOCAR LADIA, RMT

[TRANS] UNIT II: RISK MANAGEMENT AND SAFETY IN THE LABORATORY

IX Control of Biological Substances Hazardous to Health

OUTLINE and the Environment
I Introduction A Handling
II Instrumentation in Histotechnology (Part 1) B Disposal of Biohazardous Waste
A Microscope X Hazards and Handling of Common Chemicals
i Multihead Microscope A Acetone
B Rotary Microtome B Aliphatic Hydrocarbon Clearing Agent
C Floatation Bath C Acetic Acid
D Cryostat D Aluminum Sulfate
E Autotechnicon E Ammonium hydroxide
i Chemical Order F Aniline
F Automated Cover Slipper
G Celloidin (Stabilized Nitrocellulose)
G Automated Stainer
H Embedding Center H Chloroform
III Instrumentation in Histotechnology (Part 2) I Chromic Acid
IV The Care and Use of a Microscope J Diaminobenzidine (DAB).
A Parts of the Microscope K Dimethylformadide
B Parts of the Lens System L Dioxane
C Magnification and Calibration M Dyes
D Importance of Numerical Aperture N Ethanol
E Resolving Power
O Ether
F Daily Care and Maintenance of the Microscope
V Types of Hazards P Ethidium bromide
A Biohazard Q Ethylene glycol ethers
B Irritants R Formaldehyde
C Corrosive Chemical S Formic Acid
D Sensitizers T Glutaraldehyde
E Carcinogens U Hydrochloric acid
F Toxic Materials V Hydrogen Peroxide
VI Physical Hazards
W Hydroquinone
A Combustibles
B Flammables X Iodine
C Explosive Y Isopentane
D Oxidizers Z Isopropanol
VII Proper Waste Disposal AA Limonene
VIII Control of Chemicals Hazardous to Health and the BB Mercuric Chloride
Environment CC Methanol
A Personal Hygiene Practices DD Nitric Acid
B Labeling
EE Nitrogen (Liquid)
i Minimum Information
C Warning Signs FF Osmium Tetroxide
D Protective Equipment GG Oxalic Acid
E Ventilation HH Periodic Acid
F Hazard Symbols II Phenol
i Health Hazard JJ Picric Acid
ii Flammable KK Potassium Permanganate
iii Exclamation Mark LL Propylene Glycol
iv Gas Cylinder
MM Silver Salts
v Corrosive
vi Explosive NN Sodium Azide
vii Flame Over Circle OO Sodium Hypochlorite (Liquid, Chlorine Bleach)
viii Poison PP Sodium Iodate
ix Environmental Hazard QQ Sulfuric Acid
G First Aid RR Toluene/Xylene
H Radiation SS Zinc Chloride
I Storage of Hazardous Chemical XI Risk Management

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INTRODUCTION  Stage: where slides are placed

 Risk management pertains to personal as well as o Clipped by Stage clips
environmental health and safety and the first step in risk  Microscope objectives
management is to identify all hazards in and emanating o Scanner (4x), Low Power (10x), High Power (40x) and
from the laboratory. Oil Immersion (100x)
 Electrical, mechanical and biological hazards must also be  Differentiated by bands
included.  Scanner (Red)
 Obsolete chemicals should be routinely disposed of.  Low Power (Yellow)
o There are chemicals that when stored for a long period  High Power (Blue)
of time can become explosive  Oil Immersion (White)
 Standard operating procedures must be detailed to
include control hazardous substances, risk assessments, MULTIHEAD MICROSCOPE
and other health and safety information relevant to handling
specimens.
o Read SOP before entering laboratory
o We are dealing with chemicals, infectious agents so,
we must know how to handle such
 Equipment malfunction due to poor maintenance and poor
quality reagents in poor processing of tissues or inaccurate
staining results.
o Equipment are our friends and must be taken care of
o Once there is equipment malfunction, processes are
delayed
 It is everyone’s responsibility to minimize risks
associated with day to day activity by using safety guards
and checking the quality of reagents.  Microscope used in histopathology
o MT is responsible to check everything needed o Usually used by pathologists
 Poor quality can affect the subsequent steps for o Main microscope at the center
the laboratory test performed  Where slides are placed
 Poor reagents for histology may result to soft  Whatever is viewed here is also viewed on both
tissue specimen which will not be good sides
o For multi learning
INSTRUMENTATION IN HISTOTECHNOLOGY
(PART 1) ROTARY MICROTOME
 Care, appropriate use and maintenance of equipment are
essential quality control in histotechnology.
 The major pieces of equipment for any surgical pathology
are:
o Microscope
o Microtome (rotary): most common type of microtome
o Cryostat: cold
 Used for frozen sections (needing immediate
diagnosis)
o Autotechnicon
 Multitasks since it does many processes (fixation,
dehydration, clearing and infiltration)
o Automated coverslipper: automated system how to  Image on the other side shows a manual and automated
mount rotary microtome
o Automated H and E stainer o Difference of automated to manual: buttons
o Embedding Center
o Floatation Bath

MICROSCOPE

 Block holder: push it forward and clip the tissue block

 Lever: used for trimming
o Trimming: expose the tissue in the block. During
embedding, the first layer is paraffin, so we need to
remove first the paraffin for us to see the tissue.
o For laterality
 Safety lock for blade

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CRYOSTAT

 Lever for angle: increase (left), decrease (right)

o Angle used is usually 5 degrees
 Flywheel: clockwise movement and the block holder will
move up and down

 Rotary microtome inside a cold chamber

 Smaller flywheel
o Clockwise, the movement is forward  Inside the cold chamber
 Towards the blade  Environment: -5 to -30 degrees Celsius
o Counterclockwise, the movement is backward.
 Tissue ribbon: paraffin that contains the tissue section
o Must be floated at the floatation bath
 Thickness adjustment knob: it is set on micra (the thinness)
o Usually 2-3 micra.
 50 degrees Celsius temperature of water bath
o For the spreading of the ribbon
o Water bath also known as the floatation bath
 Afterwards, proper labeling is done.
o The label of the block is the label of the slide.
 Fishing out/Fish out: transfer of ribbon to the glass slide.
o Technique
 Tissue section must be centered
 1st image: Cryotome Disc
 Tissue sections must be placed to cold water going to hot
 2nd image: weighing compressor
water
o Compress at cryotome disc
o Tissue sections must be spread out  5th image (2nd at the lower part): image of the appearance
 To avoid folds and so we can see properly in the of the tissue specimen
microscope o Tissue specimen is surrounded by a white material
(seen on the 4th image(first at the lower part))
FLOATATION BATH  Tissue freezing medium
 Last image: appearance of a tissue with tissue freezing
medium
o Specimen is placed on a disc then poured with
medium, and once it hardened, it will be as seen in the
last image

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 Cryotome Disc is placed in the hole and lock it by rotating

clockwise
 As long as safety lock is lowered down, it is safe and you
will not be bruised.
o Safety precaution

 There is no floatation bath here

o Difference: in the earlier image
 We put the ribbon on floatation bath, but here, we
 Clockwise rotation: up and down movement just compress it on the slidee

 Difference with the rotary microtome

o It has button
o If the movement of the rotary is on the left side (smaller
fly wheel), clockwise forward, counterclockwise
backward
 For the image above, it has only button, press it
down and the specimen will move forward, press
it upward and the specimen will move backward
 Same principle with rotary microtome
 Staining
 End result
 Specimen is at the center.
o One tissue section
 There is three because once the other sections are
destroyed. You have an extra specimen.
o For safety (extra tissue section)

AUTOTECHNICON

 Tissue ribbon making

o Get the tissue ribbon

 The pathologist have finished grossing and so, the next

processes is given to medical technologist

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o MT starts to work  Same principle with cover slipping except that it has robotic
o For their convenience, the use autotechnicon arm
 Four processes all in one: fixation, dehydration, clearing o Slide is placed in the glass slide drawer and then it will
and ISA PA enter and there are cover slip magazines
o We have a jar and the container to put our tissue o When the slide reaches the bottle for coverslip
cassettes mountant, it will just drop mountant and then the robotic
 Usually called basket and is usually put in jar arm will get a coverslip and will compress it at the glass
o Can accommodate up to 50 tissue cassettes. slide
 If it is full, you will place it on the first jar o Mountant is controlled
 Depending on the brand  Compared in manual that has to have appropriate
technique
CHEMICAL ORDER
 First: Fixative: 10% buffered formalin AUTOMATED STAINER
 Second: Dehydration: 95% alcohol
o Depends if acetone is included (dependent on labs)
 Third: Clearing: Xylene
o Most common
o If a laboratory includes acetone, then xylene is the
fourth jar
 Fourth: Infiltration/impregnation: paraffin wax
 Autotechnicon must be programmed
o The four processes last for 8 hours
o Avoid mistakes and always check reagents (if passed
QC)

 Start with Xylene, Absolute Alcohol, then 95% alcohol,

wash with water and hematoxylin.
 Hematoxylin will depend on the Quality Control.
o Dependent on how many minutes it is dipped
o The more concentrated, the lesser the time is.
 The automated stainer has robotic arm
o It will first dip with xylene
 Dependent on the program (e.g. 10 dips) and will
 Ex. 10% buffered formalin on the first jar. However, the move to the next steps stated above
process depends on the program you had done (duration of  Move right
the process)
 We need agitation for the tissue section to absorb the
chemical
EMBEDDING CENTER
 Manual method: dipping into formalin for 5 minutes
AUTOMATED COVER SLIPPER

 1: paraffin dispenser
 2 and 3: can be interchangeable
 2: cold chamber (0 to -10 degrees Celsius)
o Where tissue sections will be frozen
 3: warm temperature (usually 50 degrees Celsius)
 Once paraffin wax is frozen, they will stick together. So that
warm water is needed for easier removal of those frozen
paraffin wax due to infiltration (since it is the preceded step)

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PARTS OF THE LENS SYSTEM

 Nosepiece: located at the end of the body tube for holding
objectives
 Objectives: consists of system of lenses located at the end
of the body tube that are held in place by nosepiece and is
closer to the slide under examination.
 Eyepiece: ocular, nearest to the eye.
 Focal length: the distance between outer lens of
objective and the cover glass of the slide under
examination.

 Usually, processes take about 4 days, depending on the

specimen submitted.
o Depends on specimen volume
 Most of the processes are done by MT

INSTRUMENTATION IN HISTOTECHNOLOGY
(PART 2)
 It is imperative that the laboratory maintain a current file
for every piece of equipment in the laboratory. This file
contains the following information:
o Name, manufacturer, model number and serial
number.
o Record of preventive maintenance performed, as
prescribed by the manufacturer
 Always check
 Especially in histopathology, once an equipment is
destroyed, all processes will be delayed as well
o Records of service call and repairs performed.
o Copy of operating manual.  Focal length: important part as well

THE CARE AND USE OF A MICROSCOPE

 The microscope is an instrument that enlarges images and
allows the visualization of morphologic cellular details
that are too small to be seen by the unaided eye.
 Usually, microscope in histopathology section is used for
quality control and reading of the pathologists

PARTS OF THE MICROSCOPE

 Base: provides support
 Arm: supports and holds the magnifying and adjustment
system  Objectives
 Stage: flat platform where the slide is placed for  No immersion objective is done in specimens that has glass
examination. slide
 Substage: located directly under the stage and hold the o Pap smears: needs oil immersion objectives
condenser and diaphragm  If we start with High Power Field, we may destroy the lenses

MAGNIFICATION AND CALIBRATION

 Magnification is the process that increase 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 160nm.
o Objective × eyepiece = total magnification
 If the microscope is fitted with a draw tube, the body tube
length will be increased, and the body tube magnification
will also be correspondingly increased.
o Objective × eyepiece × working tube length = total
magnification
o 40 × 10 × 180/160 =450x

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 Draw tube is added to the normal tube length
divided by the normal tube length
IMPORTANCE OF NUMERICAL APERTURE
 The numerical Aperture (NA) or light gathering capacity of
lens governs the resolution and wavelength of light
employed. It measures resolving power of the objective,
is engraved on each objective, and is used to rate
objectives.
RESOLVING POWER
 The resolving power of an objective is its ability to allow the
examiner to see two particular points that are close
together, as separate and distinct.
o E.g. in one slide, there is bacilli and cocci (magkadikit),
you need to differentiate them to determine that
resolving power is okay
 Resolving power is measured by the distance between two
linear dots, or the number of lines to an inch that can be
seen separate from each other.
 The ability of an optical system to form distinguishable
images of objects separated by small angular distances
 The ability of a lens to distinguish small objects in close
proximity
DAILY CARE AND MAINTENANCE OF THE
MICROSCOPE
 A Skilled individual should perform preventive
maintenance.
 Internal prisms should never be touched.
 Lenses should be wiped daily as necessary with fresh lens
paper.
o Not an ordinary paper because if it is ordinary paper,
magagasgasan yung lenses/objective
 Immersion oil should be removed immediately after use.
 Oil can be removed using a lens paper damped with xylene.
o Once oil is hardened, it is difficult to be removed
 Always clean the objective
TYPES OF HAZARDS
BIOHAZARD
 Can be infectious agents themselves or items contaminated
with them
 Anything that can cause disease in humans, regardless of
its source.
IRRITANTS
 Chemicals that cause reversible inflammatory effects at
the site of contact with living tissue.
o Pwede pang maagapan
 Most often, eyes, skin and respiratory passages are
affected.
DELA CRUZ, GALANTO
CORROSIVE CHEMICAL
 Present both physical and health hazards
 When exposed to living tissue destruction or irreversible
alteration occur
o Causes tissue destruction
SENSITIZERS
 Cause allergic reactions in a substantial proportion of
exposed subjects.
 True sensitizers are very serious hazards, because
sensitization lasts for life and only gets worse with
subsequent exposure.
 Example-formaldehyde
o Once this builds up in the body, complications occur,
especially, carcinogens
CARCINOGENS
 Substance that induces cancer
 Chloroform, chromic acid, dioxane, formaldehyde, nickel
chloride and potassium dichromate.
 Dyes-auramine O, basic fuchsin, and any dye derived from
benzidine (congo red)
TOXIC MATERIALS
 Capable of causing death by ingestion, skin contact or
inhalation, at certain specified concentrations.
 Methanol, chromic acid, osmium tetroxide and uranyl
nitrate.
PHYSICAL HAZARDS
COMBUSTIBLES
 Have flash points at or above a specified temperature at
which vapors will ignite in the presence of ignition source
under carefully defined conditions using specified test
equipment.
 It is a guide to the likelihood of which vapors might ignite
under real workplace conditions.
o Especially alcohols
 They should not be placed close to one another
(others also include those ignition sources,
chemicals)
FLAMMABLES
 Have flash points below the temperature
o But require special designated storage rooms, cabinets
and containers, to control and prevent vapors from
building up around electrical devices that spark.
EXPLOSIVE
 Rare in histology
o Sometimes, picric acid is also included in
histopathology section
 Primary example is picric acid certain silver solutions may
become explosive upon aging, which is why they should
never be stored after use.
 Explosions may occur simply by shaking.
OXIDIZERS
 Initiate or promote combustion in other materials
o Combine with other chemicals for it to become harmful
 Harmless by themselves, they may present a serious fire
risk when in contact with suitable substances.
 Sodium iodate - mild oxidizers
 Mercuric oxide and chromic acid - serious oxidizers
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PROPER WASTE DISPOSAL HAZARD SYMBOLS

 Observe proper waste disposal using the color-coded
waste bags. Properly dispose of contaminated laboratory HEALTH HAZARD
supplies to designated disposable containers.  A cancer-causing agent (carcinogen) or substance with
o MTI (MedTech Interns) should not just throw tissue respiratory, reproductive or organ toxicity that causes
sections in trash bin damage over time (a chronic, or long-term, health hazard)
 Special procedures must be done o There are recorded cases that males may become
 Sharps and needles are thrown in puncture-proof infertile due to chemicals
container.
 Clean and disinfect any spillage by using 1:10 dilution of
bleach or sodium hypochlorite.
 Cordon the area when large spillage is done on the floor to
avoid spread of contamination. Clean and disinfect with
1:10 dilution of bleach or sodium hypochlorite.

Table No. 2-1

COLOR OF TYPE OF WASTE
CONTAINER/BAG FLAMMABLE
BLACK Non-infectious dry waste
GREEN Non-infectious wet waste
(kitchen, dietary, etc.)
YELLOW Infectious and Pathological
waste
YELLOW WITH BLACK Chemical waste including those
BAND with heavy metals
ORANGE Radioactive waste (nuclear med
laboratory)
RED Sharps and pressurized
containers
 Yellow and Yellow with Black Bands are usually used in  Very common (also in gasoline stores)
histopathology section
EXCLAMATION MARK
CONTROL OF CHEMICALS HAZARDOUS TO  An immediate skin, eye or respiratory tract irritant, or
HEALTH AND THE ENVIRONMENT narcotic

PERSONAL HYGIENE PRACTICES

 There must be no eating, drinking or smoking in the lab.
 Wash hands frequently
 Solutions must never be pipette by mouth.

LABELING

MINIMAL INFORMATION
 Includes: GAS CYLINDER
o Chemical name and if a mixture, name of all  Gases stored under pressure, such as ammonia or liquid
ingredients nitrogen
o Manufacturer's name and address
o Date of purchase and name
o Expiration date, if known
o Hazard warnings and safety precautions.

WARNING SIGNS
 Different guidelines

PROTECTIVE EQUIPMENT CORROSIVE

 General guidelines for clothing suitable for laboratory work
should be considered before protective equipment.
 Aprons, goggles, gloves and respirators are the personal
protective equipment (PPE) most likely to be used.

VENTILATION
 Ensuring proper airflow through laboratory is the first
critical step in improving working conditions.
o In histopathology section, the MTs will not enter the
section if exhaust fan is not yet opened
 Closed exhaust fan will let MTs exposed to vapor
 Image showing test tube
of formalin

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o Poured at hands and on surfaces  Dangerous liquids are best stored below countertop
height to minimize the risk of bodily exposure in case a
EXPLOSIVE bottle is dropped and broken.
o Ex. in the lower area of cabinet
 Specialized acid cabinets are designed to contain the
fumes emanating from most containers of strong mineral
acids.
 Transporting hazardous materials from storage to work
areas can be risky

CONTROL OF BIOLOGICAL SUBSTANCES

HAZARDOUS TO HEALTH AND THE ENVIRONMENT

HANDLING
FLAME OVER CIRCLE  Three potential routes of exposure
 Oxidizers o Inhalation of aerosols
o Contact with non-intact skin
o Contact with mucous membrane (eyes, nose and
mouth)
 Knowing how infectious agents can reach you is the
foundation of protecting yourself and your co-worker.
o Need to know infections agents and chemicals to
protect yourself
 Fresh specimens of human origin must always be
considered potentially infectious.
o Especially fresh tissues and body fluids
 The first and most obvious source of biological risk is with
fresh tissue and body fluids.
POISON
 Grossing carries the highest risk of all histological activities.
 Skull and crossbones  Fixed specimen have a much-reduced risk because nearly
all infectious agents are readily deactivated by histological
fixation.

DISPOSAL OF BIOHAZARDOUS WASTE

 Should be incinerated on site or hauled away
 Segregate waste
 Fixed wet specimen

HAZARDS AND HANDLING OF COMMON

ENVIRONMENTAL HAZARD CHEMICALS

ACETONE
 Highly flammable and very volatile
 Can be narcotic in high concentration.
 Such chemical is monitored by PDEA
 Also used as a polish remover

FIRST AID
 With laboratory chemicals, the most common accidents
requiring first aid are ingestion, eye contact and extensive
skin contact.
 Should have basic training in dealing with these situations
 Emergency eyewash stations
o NSS is used if the condition is worsened ALIPHATIC HYDROCARBON CLEARING AGENT
 Go to ER already  Very low toxicity, nonirritating, and non-sensitizing to
 Emergency showers normal human skin.
 Combustible or flammable
RADIATION  Recycle by fractional distillation
 If radioactive substances are handled, a qualified radiation
safety officer must oversee all aspects of the project,
ACETIC ACID
including disposal
 Used as fixative
 Workers should wear dosimeters
 Irritating to respiratory system, corrosive to most metals
o Used in nuclear medicine laboratory
o Measures the radiation that is absorbed by the body  Use a chemical fume hood, nitrile gloves, goggles and
impermeable apron when dispensing concentrated acid.
STORAGE OF HAZARDOUS CHEMICAL
 Can be safely stored in conventional cupboards.

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 Always add acid to water never water to acid, to avoid
severe splattering/reaction
 Do not mix concentrated (glacial) acetic acid with chromic
acid, nitric acid or sodium /potassium hydroxide.
o These are decalcifying agent
 If specimens are hard, e.g. bones, we must not
use fixative as acetic acid
ALUMINUM SULFATE
 Not dangerous in laboratory except as eye irritants
AMMONIUM HYDROXIDE
 Very dangerous
 Severe irritant to the skin, eyes, respiratory tract
 Target organ effects on respiratory system (fibrosis
and edema)
 Store away from acids
o Must be separated from acids, especially decalcifying
agents
 Especially when vapors are generated
 Do not mix with formaldehyde as this generates heat
and toxic vapors
 Spills of 500mL or more warrant evacuation in the room.
o Ammonium hydroxide is used to create ammonia
water.
o Ammonia water: used as a bluing agent
o Bluing agent: intensifies the staining of hematoxylin.
 The procedure only involves mixing it with water.
ANILINE
 Very dangerous reagent, which should not be used if
possible.
 Moderate skin and severe eye irritant, sensitizers, toxic by
skin absorption, carcinogen,
 Excessive exposure may cause drowsiness, headache,
nausea, and blue discoloration of the extremities.
o It is also used as a clearing agent, but Xylene is the
most common.
DELA CRUZ, GALANTO
CELLOIDIN (STABILIZED NITROCELLULOSE)
 Harmless as health hazard but dangerously flammable as
solid.
 Solutions usually contain highly flammable ether and
alcohol.
o Used for embedding and infiltration.
 The most common used in these steps is paraffin
wax, but Celloidin is just an alternative one.
o Although harmless, it is dangerously flammable as
solid.
 Has ether and alcohol
CHLOROFORM
 Used in clearing
 Toxic by inhalation and ingestion
 Overexposure can cause disorientation, unconsciousness
and death
 Target organ effects on liver, reproductive, fetal, CNS,
blood and GIT
 Carcinogenic
 One of the most dangerous and difficult chemicals in
histology because workers in most laboratories simple
cannot receive adequate protection from vapors and skin
contact.
 Do not burn
 Do not evaporate solvent to the atmosphere (always
closed)
 Avoid all use
o Chloroform is used in clearing.
o It is very toxic since its target organ is the CNS, blood,
and GIT.
CHROMIC ACID
 Highly toxic with target organ effects on kidney, corrosive
to skin and mucous membranes, carcinogenic.
 Strong oxidizer
 Chromium is a very serious environment toxin.
 Give this chemical high priority for complete elimination
from your lab.
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o This is not usually used in the laboratory here in the

Philippines.
DYES
 All should be handled with due to caution when in the
powder state, but liquids pose little risk except through
skin contact and ingestion.
 Usually used in stains

o Toxic and a strong oxidizer.

o Mostly, this is not use in the laboratory.

DIAMINOBENZIDINE (DAB)
 Human carcinogen
 Solutions pose little health risk under normal conditions of
use
 Do not use chlorine bleach as the reaction, products
remain mutagenic
ETHANOL
 Skin and eye irritant
 Flammable liquid
 Recycle via distillation

o Used in immunohistochemistry.
o It appears color brown in the nucleus and cytoplasm.
o It is risky to use since it is a cancer inducer chemical.
ETHER
DIMETHYLFORMADIDE  Mild to moderate skin and eye irritant
 Eye, nose and skin irritant. Nay cause nausea  Overexposure to vapors can produce disorientation,
 May be reproductive toxin unconsciousness or death.
 Combustible liquid  Target organ effect on nervous system following inhalation
or skin absorption.
 Dangerously flammable liquid that forms explosive
peroxides.
 Placed in metal container

DIOXANE
 Skin and eye irritant, overexposure may cause corneal
ulceration
 Flammable liquid that develops explosive properties after
a year. Do not recycle.
 Avoid all use of this chemical.

ETHIDIUM BROMIDE
 May be harmful by ingestion, inhalation, or absorption
through the skin
 Chronic exposure may cause alteration of genetic
material
o Especially important in pregnant workers in histopath
lab, usually, we do not have pregnant workers and
those with cancer history in such lab because of these
chemicals

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GLUTARALDEHYDE
 Severe eye and skin irritant

ETHYLENE GLYCOL ETHERS

 Toxic by inhalation, skin contact and ingestion, with target
organ effects involving reproductive, fetal, urinary, and
blood systems
 Combustible liquids
 Avoid all use, substituting propylene-based glycol ethers.

o Fixative and an irritant.

HYDROCHLORIC ACID
 Strong irritant to skin, eyes, and respiratory system
 Concentrated acid is particularly dangerous because it
fumes.
 Target organ effects via inhalation on respiratory,
reproductive and fetal system.

o Usually used in the frozen section.

o But now, it is safer to use propylene-based glycol
ethers.

FORMALDEHYDE
 Severe and eye irritant
 Most serious hazard for most laboratory workers
 Toxic by inhalation and ingestion
 Target organ effects or respiratory system
 Carcinogen HYDROGEN PEROXIDE
 Corrosive to most metals  Solutions less than 5% is harmless
 Exposure of the skin during grossing is the greatest risk in  Concentrated solutions are hazardous
a well-ventilated lab.

o Hydrogen peroxide: used in bacteriology and used in

Catalase Test for S. aureus.

o Grossing: specimen is taken from the formalin. HYDROQUINONE

 Irritant capable of causing dermatitis and corneal
FORMIC ACID ulceration
 Used as decalcifying agent  May cause dizziness, sense of suffocation, vomiting,
 Irritant headache, cyanosis, delirium and collapse
 Mild skin and severe eye irritant  Avoid contact with sodium hydroxide
 Corrosive to metal
 Use chemical fume hood

DELA CRUZ, GALANTO CSU BS MLS 3B 12

 TRANS: RISK MANAGEMENT AND SAFETY IN THE LABORATORY

IODINE
 Strong irritant and corrosive to eyes, skin and respiratory
METHANOL
system
 Moderate skin and eye irritant
 Dermal sensitizers
 May cause blindness or death
 Histological solutions are essentially harmless except if
 Flammable and rather volatile
ingested
 Recycle

ISOPENTANE
 Excessive exposure to vapors causes irritation of
respiratory tract, cough, mild depression and irregularly
beat.
 Extremely flammable and highly volatile NITRIC ACID
 Commonly used decalcifying agent
ISOPROPANOL o Mixed with formalin to create a good decalcifying agent
 Toxic by ingestion  Corrosive to skin, mucous membranes and most metals
 Flammable liquid  Toxic by inhalation
 Recycle by distillation

LIMONENE
 Dangerous sensitizers when handled in histology
 Safe food additives
 Cannot be recycled back

o Do not put concentrated nitric acid on tissues.

 If tissue is dipped on the acid, it will soften and turn
yellow.
 Therefore, once cut, the tissue section will not be
o Used as a clearing agent but a dangerous sensitizer.
complete, hence it is filled with holes because of
o It is less toxic than xylene, but xylene is preferred
the reaction / effect.
 Because xylene is better on tissue absorption
 Limonene is also expensive
NITROGEN (LIQUID)
 Can cause frost bite or thermal burn
MERCURIC CHLORIDE
 Excessive inhalation may cause dizziness, loss or
 Severe eye irritant, target organ effects on reproductive,
consciousness or death from asphyxia
urogenital, respiratory, GIT, and fecal system.
 Usually used in frozen sections
 Severe environmental hazards
 Corrosive to metals
 Legitimate to disposal of mercury containing waste is
difficult and very expensive.

DELA CRUZ, GALANTO CSU BS MLS 3B 13

 TRANS: RISK MANAGEMENT AND SAFETY IN THE LABORATORY

PICRIC ACID
 Explosive when dry or when combined with metal and
metallic salts
 Should not be disposed by pouring down the drain since
they may form explosive picrates with metal pipes.
 Jars and cap threads containing picric acid should always
be wiped with damp towel to prevent the substance from
drying.
 Toxic when absorbed through the skin

OSMIUM TEROXIDE
 Corrosive to eyes and mucous membranes
 Vapors are extremely toxic to reproductive, sensory and
respiratory systems.

o When dried, it will be explosive.

POTASSIUM PERMANGANATE
 Cause irritation of skin and eyes
 Strong oxidant, should not be mixed with acetic acid,
ammonium hydroxide, ethanol, ethylene glycol,
OXALIC ACID glycerol, hydrochloric acid, hydrogen peroxide or
 Safe when diluted sulfuric acid.
 When concentrated, it is corrosive and causes severe o Causes reactions to such
burns of the eyes, skin and mucous membranes

o It is diluted because it is harmful when it is on the

powdered form.
o Most difficult chemical to handle.
PERIODIC ACID
 Relatively safe PROPYLENE GLYCOL
 Less toxic, substitute for ethylene-based ethers
PHENOL
 Readily absorbed through the skin, and may cause SILVER SALTS
increased heart rate, convulsion or death, or may burn eyes  Safe when used as fresh solutions, but can be explosive
and skin. when solutions became old
 Combustible  Serious environmental hazard and should not be
discarded down the drain.

DELA CRUZ, GALANTO CSU BS MLS 3B 14

 TRANS: RISK MANAGEMENT AND SAFETY IN THE LABORATORY

SODIUM AZIDE
 Very toxic and may be fatal when swallowed or absorbed
through the skin, or when mixed with acids
 It explodes when placed in contact with metals and
should not be discarded down the drain.

SODIUM HYPOCHLORITE (LIQUID, CHLORINE

BLEACH)
 Strong oxidant, eye irritant and corrosive to most metals
 Do not mix bleach with formaldehyde or o Commonly used clearing agent.
diaminobenzidine (DAB) o Xylene can melt rubber gloves.

ZINC CHLORIDE
 Corrosive to most metals, including stainless steel.
 Should not be used in tissue processor.
 Skin and eye irritant
 Can cause severe GI problems if ingested.

SODIUM IODATE
 May be used to replace mercuric oxide when reconstituting
Harris hematoxylin.
 Colored Pink

o Note: do not use on Autotechnicon, because it is

corrosive on metals, even though it is stainless.

RISK MANAGEMENT
 Identify and evaluate hazards
 Plan to minimize the risk
 Implement the plan
 Design SOP for working with hazards
 Train personnel
SULFURIC ACID  Periodic review
 Strong irritant to the skin, eyes and respiratory system  Record keeping
 Corrosive to metals
 Dilute solutions pose no risk, while concentrated acid
produces fumes that are dangerous to health and require REFERENCES
the use of fume hood, apron, goggles and gloves.
Notes from the discussion by Prof. Jocar Ladia, RMT

Things may not be smooth sailing, but being determined and

having that big leap of faith in you, things will become possible.
The first thing you must be proud of is you being able to finish
this reviewer.

Another introductory part, may this help you aspire more, work
more and do more of what you think you can do because you
must not be limited by what others perceives about you. Be
TOLUENE/XYLENE yourself, work and believe you have the power to do things
 Skin and eye irritant because you are who you are!
 Repeated exposure can cause impaired memory, poor
coordination, mood swings and permanent nerve God Bless soon to be MTI and fRMT
damage

DELA CRUZ, GALANTO CSU BS MLS 3B 15