Histopath Lab Rules and Regulations – wear n95 mask
Eating, drinking and unnecessary personal
and goggles to avoid irritation
1. Always disinfect your working area before and
after performing each procedure
2. Always put on proper paraphernalia when
dealing with human tissues, secretions as well
as chemicals
3. To protect your clothing, always wear your lab
gown. However, gowns must not be worn
outside the lab
4. One of the most common accidents in the lab.
Involves the cutting of one’s finger or hand in
microtome knives. To avoid this, always learn
the basics of the microtome’s operation before
using it
a. Microtome – core equipment used in
lab
5. Most of the chemicals used in a histopath lab
can be handled safely, however, some o the
chemicals are potentially hazardous. Below are
some of the chemicals ad their nature
a. Ammonium hydroxide – must be stored
away from acid (formaldehyde) should
not be mixed with formaldehyde bc can
generate heat
b. Formaldehyde – acidic substance, can
generate heat and toxic vapors, very
toxic when inhaled much more when
ingested. Both carcinogenic and
corrosive. Prolonged exposure develop
into cancer cells
c. Xylene – form of irritant, can irrritae te
skina dneyes when ingested it can be
toxic in the body. Repeated exposure
can lead to contribution of dysfunction
of CNS (impaired memory, poor
coordination, mood swings, permanent
nerve damage (use is restricted in
unimportant tests) used in mounting
technique
6. Practice proper hand washing. Do wash your
hands before and after each lab technique
(most esp when handling tissues)
7. If you are injured, notify your instructor
immediately (if hospi notify chief medtech)
8.
grooming is STRICTLY NOT ALLOWED inside the
laboratory (lounge room pwede magkain mag
arte to avoid exposure to diff substances)
ROUTINE PROCEDEURE IN HISTOPATH
1. Receiving/numbering
 When going to receive specimen must
check the label
 When receiving the specimen always
check if the specimen is completely
soaked with formalin to preserve
i. Para di mawala integrity and di
mahirap mag identify
 Sample must e soaked in fixative to
maintain integrity
 Check label
2. Labelling/Filing
 After receiving the sample record the
data in the assigned logbook
 Always check the label if it includes the
data of the patient such as name of the
patient, what type of organ, name of
the doctor who operated and date of
operation to protect patients from
adverse errors made due to improperly
labelled specimen, what is sample, age
and sex
 Record data in assigned logbook
 Per spx diff logbook and have column
for result
 Always right not computerized so that
may back up incase masira computer
3. Gross Examination
 Pathologist must perform this
 Characteristics that are noted are size,
color, consistency of tissue or texture
(usually performed)
 Ask for thickness it must be 5mm to
allow observation of tissues under
microscope
 If sample is taken from lungs (lung
tissue) (thickness 1-2cm) it looks
gelatinous so it is prone to shrinkage
when fixation and other process starts
  Pathologist will get representation from
the are for example if there is
discoloration on this part then doon
magkuha na portion ang patho
 Medetch is the assigstant (note evrthing
 Each discoloration are noted and spx
 Ideal thickness is 5mm or millimeter
 Consistency of lung inside is gelatinous
(ideal thickness is 1-2cm)
 1-2cm dapat if ever magshrink may
naiwan pa
4. Fixation
 The purpose is to preserve tissue and
should be placed in fixative after
removal in body para di malata
 We have a special room for storage of
organs because there is a tendency na
magkaso ang patient for retrieval
purposes
 Formalin or formadeldyhe - #1 fixative
used
 From operating room to lab naka soak
na sa fixative
5. Decalcification
 Decalcify 24-48 hrs until it is soft
 Performed in order to soften a harden
tissue
 Hard ang bone bcos of calcium salts
present
 Ex: bones, teeth, cartilage
 Will remove the calcium salts and lime
deposits
 If di maglambot masira ang microtome
6. Dehydration
 Intracellular and extracellular water is
being removed from tissues after
fixation process to remove formalin
 Done after fixation
 Tissue will be dehydrated
7. Clearing
 The process is the excess dehydrating
agent is being removed from tissue and
then replace by a substance that will
remove/dissolve infiltrating medium
 Clear dehydrating agents
8. Infiltration
 Interstitial tissues and cavities are being
impregnated with the medium (paraffin
wax for infiltration) in order to allow the
cutting of the sample
9. Embedding
 The impregnated tissues that is being
processed in infiltration, it will to
solidify to allow the setting of sample
 Impregnating tissue will be put in a case
in order to solidify the medium
10. Trimming
 Naa man jud excess amount of wax na
nag surround sa tissues so we are going
to trim the excess wax in order to
expose the tissue itself and after we are
going to cut into section into very thin
slices using microtome machine after
achieving thin slice
 Excess wax will be removed or cut off in
order to expose the tissue
11. Sectioning
 Microtome cut sections ito very thin
slices using microtome
 Must have the capacity to slice into very
very thin section
 Thin section is needed in order for
patho to check the cells para di mag
overlap
 Di maview ang characteristics
 The thinner the better
12. Staining
 Process of adding color to tissue slices
to enhance visualization and
differentiation of cellular structures in
the microscope
 Para ma appreciate anong appearance

13. Mounting
 Place a single drop of medium (xylene)
on top of sample (can preserve sample
and for better viewing) and cover it with
coverslip to preserve the sample bc we
need to store it up to 10 years
Serial number – accompanied by letter that signifies the
type of spx transmitted (autopsy – A-102) if from
surgery or biopsy (S-102)
- Given in histopath lab\\
RESPONSIBILITY OF A LAB TECHNICIAN
1. SPECIMEN PRESERVATION (even formalin, mix
with water)
2. SPECIMEN LABELING, LOGGING AND
IDENTIFICATION
3. PREPARATION OF THE SPX
4. RECORD KEEPING
Classification of chemicals
1. Biohazards
a. Infectious agent
b. Can be contaminated and our health is
pit at risk
2. Irritants
a. Can result to inflammatory effects dute
to irratation
3. Corrosive
a. Have both physical and health hazards
b. Physical – may chance to cause fire
c. Fire – result to destruction or
irreversible alteration on living tissu
4. Carcinogens
a. Result to cancerous cells
b. Ex: formalin, chloroform
c. Potassium dichromate
a. HISTOPATH personnel make sure that
label is correct
b. Owners must have training programs
c. Non-medtechs (janitor) – must be
oriented as well bc they enter the lab
4. Corrosive substances – cause injury to skin and
eyes if may direct exposure or contact
a. Possible effects on cell – encourage r
lead to cellular membrane disruption,
coagulation of proteins, or death of the
cell (usually f may ingestion of the
substance)
5. Fire and explosion hazards – we have liquid
form and dry/powder form, these are fire
hazards bc they behave capacity to explode drid
(picric acid, benzoyl peroxide, silver ions)
a. Fire distinguisher dapat anjan
6. Hazardous chemical spills and storage – lab
personnel who will clean the are mjst be trained
kung ano gamitin o gawin if may mahulog a
floor dapat may absorbent pads to remove
spillage sa floor wag maggaimit tubig kay baka
magreact sa water
a. Spill kits – to get if there is spillage. May
gloves, aprons, absorbent pads, use to
clean spillage

Hazards in hisopath lab

1. Biological or infection hazards – put tissues and

objects with contact on that rissue in biohazard
bags and hsharp for sharps
a. Tiuberculosis exposure
b. HIV, HVC, HBV
c. Handling tissue waste
2. Mechanical hazards – sharp instruments
(microtome blade, scalpel, broken glass, razors)
3. Chemical hazards – materials afety data sheet
dapat meron (inside this has all info abt
chemcical like how to handle, storage temp,
saan ilagay)
ACTIVITY NO. 1 INSTRUMENTATION a. Container – for sample
b. Molder – mold wax here for infiltration
c. Tissue cassette – after tissue is sliced
put here. Have no label normally. Put
serial # using pencil. Must be readable.
Each step in the tissue processing necessitates the use Color coding in US
of various specific components. As such, the rooms in t i. Yellow – liver tissues and renal
tissues
ii. Green – routine tissues
Components of histopath lab (fallopian tube, cervix,
gallbladder, appendix)
1. Equipment and apparatus iii. White – bone sample
a. Diamond pen, pencils – to have iv. Gray – sample is skin
permanent label on glass slide v. Pink – lymph node
b. Gross table 3. Chemicals and reagents
i. Forceps 4. Dyes
ii. Scalpel
iii. Chopping board
iv. Weighing balance
v. Ruler
vi. Orientation markers
vii. Watch glass/petri dish
viii. Tissue cassettes
ix. Saw – for bigger bones
c. Automated tissue processor (FDCI) – 12
hrs
d. Paraffin Oven – to melt paraffin, slide is
put here
e. Embedding Center – impregnation and
embedding is performed here
i. Hot first
ii. Cold next – for ghardening of
paradffin wax or tissue blcks
f. Refrigerator – 4-6 degree celsiums
g. Microtome
h. Flotation bath – after blocks are section,
put the sample here. Temp is hot konit
to allow manipulation of sample into
the slide and para magdikit sample sa
slide
i. Side dryers – can use paraffin oven if
wala ito
i. Automated stainers
ii. Microscope – pathologists are
the ones using this
2. Wares – for holding substances