HISTOPATHOLOGY

Histopathologic Techniques
College of Medical Laboratory Science | OLFU | 2024

OUTLINE
● Histopathologic ○ Embedding
Techniques ○ Blocking
○ Numbering ○ Trimming
○ Fixation ○ Sectionning
○ Dehydration ○ Staining
○ Clearing ○ Mounting
○ Impregnation ● Ferning Specimen source
1. Bilateral organs
HISTOPATHOLOGIC TECHNIQUES ○ Examples: Extremities, kidneys,
● involves different procedures that have lungs and ovaries)
been adopted for the preparation of 2. Miscellaneous
materials and tissue for microscopic ○ Examples: age, sex, ward
examination
RECEPTIONIST
12 STEPS IN HISTOPATHOLOGIC TECHNIQUES: ● Validates if the specimen is adequate or
1. Numbering 7. Blocking good for tissue processing
2. Fixation 8. Trimming ● 1st person that will receive the specimen
3. Dehydration 9. Sectioning and put it into the container
4. Clearing 10. Staining ● CONTAINER: contain fixatives
5. Wax impregnation 11. Mounting Proportional to the size of the
6. Embedding 12. Labeling specimen
Clear
“Tissue processing” steps Unbreakable
1. FIXATION: fix tissue for processing into thin Wide mouthed bottle
microscopic sections.
2. DEHYDRATION: water from wet tissues are II. FIXATION
removed by increasing concentrations of ● Most critical step in histopathological
alcohol (70% to 95% to 100%). techniques
3. CLEARING: once successfully dehydrated, ● PRIMARY AIM: preserve the morphology
dehydrating agent (alcohol) is removed and chemical constituents of the tissue.
with a clearing agent (xylene is most ● SECONDARY AIM: Protect and harden the
common) specimen for further handling
4. INFILTRATION: tissues infiltrated with an
embedding agent (paraffin).

I. NUMBERING
Basic information needed:
1. Date and time
2. Name of the patient
3. Specimen Number
○ C- Cytology specimen
○ A- Anatomical specimen
○ S- Surgical specimen
■ Example: S-09-2111

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HISTOPATHOLOGY - Histopathologic Techniques

FIXATION STEPS: TYPES OF FIXATIVES

1. specimen is placed in a liquid fixing agent (ACCORDING TO COMPOSITIONS)
(fixative) such as formaldehyde solution 1. Simple Fixative - Uses only one chemical
(formalin) for fixation
○ PURPOSE: slowly penetrate the 2. Compound Fixative - use of two or more
tissue causing chemical and chemicals for fixation
physical changes that will harden
and preserve the tissue and protect FIXATION METHODS:
it against subsequent processing 1. Acetone Fixation ● Fix cells in -20°C
steps acetone for 5-10
○ CHEMICAL: Formalin - minutes
phosphate-buffered solution - ● No permeabilization
most popular fixative for step needed
preserving tissues that will be following acetone
processed for paraffin embedding fixation.
2. Specimens should remain in fixative long 2. Methanol Fixation ● Fix cells in -20°C
enough methanol for 5-10
○ PURPOSE: (ideally) to penetrate the minutes.
tissue and then for an additional ● Permeabilization step
period in order to allow the is needed following
chemical reactions of fixation to methanol fixation.
reach equilibrium (fixation time). 3. Ethanol Fixation ● Fix cells in cooled 95%
3. Trimmed specimens are placed in suitable ethanol, 5% glacial
labeled cassettes (small perforated acetic acid for 5-10
baskets) minutes
○ PURPOSE: to segregate them from 4. Methanol-Acetone ● Fix in cooled
other specimens. Fixation methanol, 10 minutes
○ Formalin-fixed, at –20 °C.
paraffin-embedded tissues may ● Remove excess
be stored indefinitely at room methanol.
temperature, and nucleic acids ● Permeabilize with
(both DNA and RNA) may be cooled acetone for 1
recovered from them decades after minute at –20 °C.
fixation 5. Methanol-Acetone 1:1 methanol and
●
Mix Fixation acetone mixture.
EFFECTS OF FIXATIVES Make the mixture
●
1. Inhibit bacterial growth and reduce the risk fresh and fix cells at
of infections -20 C for 5-10
2. Act as mordant or accentuator - minutes.
accelerating the staining process
6. Methanol-Ethanol ● 1:1 methanol and
Mix Fixation ethanol mixture.
TYPES OF FIXATIVES
● Make the mixture
(ACCORDING TO ACTIONS)
fresh and fix cells at
1. Microanatomic Fixative - Involves small -20 C for 5-10
tissue or organ minutes.
2. Cytological Fixative - Involves body fluid or
7. Formalin FIxation ● Fix cells in 10% neutral
secretion
buffered formalin for
3. Histochemical Fixative - Involves tissue
5-10 minutes.
containing labile substances
● Rinse briefly with PBS.

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HISTOPATHOLOGY - Histopathologic Techniques

● Permeabilize with 3. Before final absolute ethanol, acetone or

0.5% Triton X-100 for isopropanol,, and chloroform or
10 minutes. trichloroethane as the transition solvent.
8. Paraformaldehyde- ● Fix in 3-4% ○ PURPOSE: ensure complete
Triton Fixation paraformaldehyde dehydration
for 10-20 minutes.
● Rinse briefly with PBS. OTHER COMMONLY USED AGENTS:
● Permeabilize with Dehydration Time (DT)
0.5% Triton X-100 for 1. Acetone ● DT: 1-2 hours
10 minutes ● colorless, cheap, rapid-acting
9. Paraformaldehyde- ● Fix in 4% 2. Dioxane ● DT: 3-24 hours
Methanol Fixation paraformaldehyde ● less tissue shrinkage
for 10-20 minutes. compared to alcohol
● Rinse briefly with PBS. ● miscible in water and paraffin;
● Permeabilize with expensive
cooled methanol for
5-10 minutes at –20 time schedule:
°C. ● (1st) pure dioxane solution 1
hour
III. DEHYDRATION ● (2nd) pure dioxane solution 1
● Utilizing chemical known as dehydrating hour
agents ● (3rd) pure dioxane solution 2
● Removing of intracellular and extracellular hours
water and fixatives in the tissue ● (1st) Paraffin wax 15 minutes
● EXAMPLES: ● (2nd) Paraffin wax 45 minutes
1. Alcohol- most commonly used ● (3rd) Paraffin wax 2 hours
2. Acetone ● Embed in mold and cool in
3. Dioxane water.
4. Tetrahydrofuran 3. Cellosolve ● rapid; combustible &
5. Cellosolve expensive
6. Triethyl phosphate ● tissue may be transferred
from water directly and stored
DEHYDRATION STEP/S: for months without distortion
1. Immerse specimens in a series of ethanol 4. Triethyl ● removes water readily
(alcohol) solutions with increasing Phosphate ● very little distortion &
concentration hardening
○ PURPOSE: to avoid excessive 5. Tetra- ● dehydrates & clears; miscible
distortion of tissue until a hydrofuran in water and paraffin
water-free tissue in alcohol is ● toxic if ingested or inhaled
reached
2. Typical dehydration sequence for Dehydrating agens for Electron Microscopy:
specimens not more than 4mm thick would 1. Ethanol - common dehydrating solvent;
be: 2. Propylene Oxide - transition fluid
○ 70% ethanol 15 min 3. Acetonitrile - substitute for propylene oxide
○ 90% ethanol 15 min
○ 100% ethanol 15 min
○ 100% ethanol 15 min
○ 100% ethanol 30 min
○ 100% ethanol 45 min

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HISTOPATHOLOGY - Histopathologic Techniques

IV. CLEARING ● expensive

● Also known de-alcoholization 8. Carbon ● similar to chloroform,
● Removal of dehydrating agent tetrachloride cheaper
● EXAMPLES: 9. Tetrahydrofuran ● superior to to others
1. Xylene- most commonly used ● perform 2 processes
2. Toluene 9. Dioxane ● time advantage
3. Chloroform ● changed 3 times within 4
4. Benzene hours
5. Cedarwood oil ● shrinkage than xylene
Other xylene ● terpenes,
CLEARING STEP/S: substitutes ● limonene,
1. Immerse tissue in one to three different ● Orange oil based clearing
xylene immersions agents,
○ PURPOSE: ethanol is gradually ● Chlorinated hydrocarbons,
replaced with xylene and when the ● Coconut oil,
tissue is embedded, the xylene will ● bleached palm oil
then be replaced by the molten
paraffin wax V. IMPREGNATION
2. A typical clearing sequence for specimens ● process whereby the clearing agent is
not more than 4mm thick would be: completely removed from the tissue and
○ Xylene 20 min replaced by a medium that will completely
○ Xylene 20 min fill all the tissue cavities.
○ Xylene 45 min ● Also known as INFILTRATION

CLEARING AGENT: PARAFFIN WAX IMPREGNATION

Clearing Time ● PARAFFIN: simplest, most common and
1. Xylene (Xylol) ● ½ to 1 hour best embedding medium used for routine
● colorless, suitable for most tissue processing
routine processing
● cost effective INFILTRATION STEP/S:
2. Toluene ● 1 to 2 hours 1. cleared tissue is infiltrated with a suitable
● preserves better, more histological wax (usually paraffin) which is
expensive & toxic liquid at 60°C and then allowed to cool to
3. Benzene ● 15 - 60 minutes 20°C in order to solidify into a consistency
● penetrates and clears that allows sections to be cut
rapidly, carcinogenic ○ PURPOSE: allow to be sectioned thin
4. Chloroform ● 6 - 24 hours enough on a microtome, forming
● slower than xylene, less ribbons that can flatten fully when
brittleness floated on a warm water bath
5. Cedarwood Oil ● 2-3 days 2. Atypical paraffin infiltration sequence of
● clear paraffin and paraffin for specimens not more than 4mm
celloidin (5-6 days) thick would be:
● CNS & cytologic studies ○ Paraffin wax 30 min
6. Aniline Oil ● not normally utilized ○ Paraffin wax 30 min
● clearing embryos, insects, ○ Paraffin wax 45 min
& very delicate specimens
7. Clove Oil ● minimum shrinkage THREE PROCESSING WAYS OF PARAFFIN WAX
● tendency to become IMPREGNATION & EMBEDDING :
adulterated, brittle TIME SCHEDULE

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1. Manual ● At least four changes of wax 1. Paraplast ● highly purified paraffin and
Processing are required at 15 minutes synthetic plastic polymers,
intervals ● 56-57°C
Fixation: 2. Embeddol ● synthetic wax substitute
● 10% Buffered Formalin 24 ● less brittle & compressible
hours ● 56-58°C
Dehydration: 3. Ester Wax ● 46-48°C
● 70% Alcohol 6 hours ● harder than paraffin, not
● 95% Alcohol 12 hours soluble in water, but is
● 100%Alcohol 2 hours soluble in 95% Ethyl Alcohol
● 100% Alcohol 1 hour 4. Water Soluble ● 38-42°C or 45-56°C
● 100%Alcohol 1 hour Waxes
Clearing: 5. Dimethyl ● thin sectioning
● Xylene or Toluene 1 hour sulphoxide
Xylene or Toluene 1 hour (DMSO)
Impregnation:
● Paraffin wax 15 minutes CELLOIDIN IMPREGNATION
● Paraffin wax 15 minutes ● CELLOIDIN (COLLODION): for large hollow
● Paraffin wax 15 minutes activities which tend to collage
● Paraffin wax 15 minutes ○ for hard and dense tissues (bones
Embedding: & teeth, whole embryo tissue)
● Paraffin wax 3 hours
2. Automatic ● automatic tissue processing Two methods for celloidin impregnation of tissue:
Processing machine (i.e., Autotechnicon) 1. Wet Celloidin Method
● only 2- 3 changes of wax ○ bones, teeth, large brain sections,
3. Vacuum ● Clear in two changes of and whole organs
Embedding xylene, for 1 hour each. ○ tissue is placed in thin celloidin
● Place the tissue in molten wax, (2-4%) for 5-7 days, transferred to
in vacuum chamber and medium celloidin (4-6%) for
make the oven airtight. another 5-7 days, drained off &
Exhaust the air slowly by poured with thick celloidin (8-12%)
means of a vacuum pump or until specimen has become
Venture suction pump until impregnated (3-5 days)
there is a negative pressure of ○ specimen is removed from the
400 to 500 mm. mercury. celloidin, transferred to an
● Leave for 15 minutes, then embedding medium containing
slowly readmit air until normal freshly poured thick celloidin and
atmospheric pressure is kept in a jar or dessicator .
reached. 2. Dry Celloidin Method
● Place the tissue in fresh wax. ○ for processing of whole eye
● Repeat steps 3 and 4. sections
● Place the tissue into fresh wax. ○ principle and procedure of this
● Repeat step 3 and leave for 30 method is similar to wet celloidin
to 45 minutes. method
● Bring to normal atmospheric ○ Gilson's mixture is added to the
pressure and embed the celloidin block before hardening, to
tissue. make the tissue transparent
SUBSTITUTE FOR PARAFFFIX WAX GELATIN IMPREGNATION
MELTING POINT ● rarely used except when dehydration is
avoided

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● for delicate specimens and frozen tissue 4. plastic

● steps: 5. aluminum foil molds
○ tissue is placed in 10% gelatin with
1% phenol for 24 hours, BASIC METHOD FOR EMBEDDING:
○ transferred to 20% gelatin with 1% 1. Open cassette to view tissue sample and
phenol for the next 12 hours, and choose a mold that best corresponds to the
finally to another fresh solution of size of the tissue. A margin of at least 2 mm
20% gelatin with 1% phenol of paraffin surrounding all sides of the
○ allowed to cool in a refrigerator tissue gives best cutting support. Discard
until impregnation and embedding cassette lid.
are completed 2. Put small amount of molten paraffin in
○ Gelatin-embedded tissues are then mold, dispensing from paraffin reservoir.
transferred to l 0% formalin for 12-24 3. Using warm forceps, transfer tissue into
hours in order to harden the tissue mold, placing cut side down, as it was
placed in the cassette.
VI. EMBEDDING 4. when the tissue is in the desired orientation,
● Also known as CASTING OR BLOCKING add the labeled tissue cassette on top of
● process by which the impregnated tissue is the mold as a backing. Press firmly.
placed into a precisely arrange position in 5. Hot paraffin is added to the mold from the
a mold containing medium which is then paraffin dispenser. Be sure there is enough
allowed to solidify paraffin to cover the face of the plastic
cassette.
● FOUR TYPES OF TISSUE IMPREGNATION: 6. Cool the top surface of the Paraplast by
1. Parrafin wax blowing gently on it. Tissues at this stage
2. Celloidin are very brittle and should be handled with
3. Gelatin care. If necessary, fill cassette with paraffin
4. Plastic while cooling, keeping the mold full until
solid.
EMBEDDING STEP/S: 7. Cool thoroughly in cold running tap water. If
1. tissue is oriented and placed in a metal you use ice water for the final cooling, you
mold that is filled with molten wax may split the block owing to too rapid
○ PURPOSE: form a solid tissue block shrinkage.
that can later be clamped into a 8. Paraplast naturally splits in the line of least
microtome for sectioning resistance-right through the tissue.
2. After orienting the section, the mold is filled 9. Paraffin should solidify in 30 minutes. When
with molten wax. the wax is completely cooled and hardened
3. The main part of the labelled cassette is (30 minutes) the paraffin block can be
placed on top of the mold and topped up easily popped out of the mold; the wax
with more wax. blocks should not stick. If the wax cracks or
4. The whole mold is placed on a cold plate to the tissues are not aligned well, simply melt
solidify. them again and start over.
5. After solidifying, the block with its attached 10. If you use plastic cups, the Paraplast block
cassette can be removed from the mold can be removed as soon as it is cooled. The
and is ready to be sectioned on a stainless steel mold should slip off easily
microtome. when cool and can be used again.

Mold types: VII. BLOCKING

1. Stainless steel ● Allows the medium to solidify to produce
2. ceramic tissue block
3. paper

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VIII. TRIMMING
● Process of removing excess wax after SECTION-CUTTING STEP/S:
embedding 1. Cutting into sections are done with a
● Can use knife/blade or heated spatula microtome
2. Thickness in which specimen is cut
IX. SECTIONNING SPECIMEN THICKNESS
● Also known as CUTTING OR MICROTOMY a. for routine Hematoxylin & 3–5 μm
● process by which processed tissue is cut Eosin (H&E)
into uniformly thin slices to facilitate b. for amyloid deposits 8–12 μm
studies under microscope c. kidney biospies 2 μm
3. Once sections are cut, they are floated on a
MICROTOME: machine or instrument used for warm water bath that helps remove
cutting sections of tissue wrinkles
● Three essential parts: 4. The cut sections are picked up on a glass
○ Block Holder - where the tissue is microscopic slide.
held in position 5. The tissue slide is drained and may be
○ Knife Carrier and Knife - for actual gently heated to evaporate the layer of
cutting of tissue sections water between the sections and the glass.
○ Pawl, Ratchet Feed Wheel and 6. When all the water is gone, the slide may be
Adjustment Screws - to line up the heated enough to melt the wax
tissue block in proper position with ○ PURPOSE: to improve adhesion.
the knife, adjusting the proper
thickness of the tissue for X. STAINING
successive sections. ● Tissue constituent are demonstrated in
sections by direct interaction with dye or
staining solution producing coloration of
the active tissue component
● HEMATOXYLIN AND EOSIN STAINING
○ Utilizes micro-anatomical studies
of tissue
○ It is a regressive staining method

STAINING PARAFFIN SECTIONS:

KINDS OF MICROTOME 1. Immerse paraffin section in a solvent (e.g.
1. Rocking Microtome most simplest and oldest xylene) two times, at 1-2 minutes duration
type of microtome each, for sections up to 10 micron thick.
2. Rotary Microtome cutting serial sections of 2. Xylene should be subsequently removed
tissue specimen with absolute alcohol, followed by
3. Sliding Microtome Most dangerous type of descending grades of alcohol
microtome ○ PURPOSE: to prevent damage and
detachment of sections.
4. Freezing Microtome urgent surgical biopsies 3. The alcohol is then finally replaced with
specimen water before actual staining of section is
5. Cryostat Microtome Permits rapid penetration performed.
(Also known as COLD of tissue biopsies for ○ Such procedure is the exact reverse
MICROTOME) surgical pathology of impregnation and may be
6. Ultrathin Microtome specimen for electron summed up by the phrase
microscope "Sections to Water".
4. After the section is cut and mounted on the
slide, it is drained and dried thoroughly to
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HISTOPATHOLOGY - Histopathologic Techniques

ensure that all moisture between the specific periods of time or

section and slide has evaporated, and that until the desired intensity of
the section is firmly attached to the slide. coloring of the different tissue
5. If an alcoholic stain is to be used, there is no elements is attained
more need to replace the alcohol with 3. Regressive ● tissue is first overstained to
water. Staining obliterate the cellular details,
6. After deparaffinization with xylene, the and the excess stain is
section is transferred to decreasing grades removed or decolorized from
of alcohol, and in such instances, the term unwanted parts of the tissue,
"Sections to Alcohol" is used, and the until the desired intensity of
staining procedure is subsequently done color is obtained
unless the tissue has been fixed in mercuric EXAMPLE:
chloride solution, in which case, the section ● Routine Hematoxylin and
is taken “to water”. Eosin (H&E) staining
7. After staining, the section is again ○ most common
dehydrated with increasing grades of method utilized for
alcohol and cleared with two changes of microanatomical
xylene to prepare the section for mounting, studies of tissues,
since most mountants are miscible in using the
xylene. regressive staining
8. The stained section may be left in xylene for ○ overstaining the
an indefinite period of time until it is finally nuclei, followed by
mounted on the slide. removal of
superfluous and
METHODS OF STAINING: excessive color of
1. Direct ● using aqueous or alcoholic the tissue
Staining dye solutions constituent by acid
● one dye is used and washed differentiation.
away after 30-60 seconds
EXAMPLE: HEMATOXYLIN AND EOSIN (H & E) Staining
● Methylene blue - blue stain ● stains thin tissue sections so that
2. Indirect ● action of dye is intensified by pathologists can visualize tissue
Staining a MORDANT which applied morphology
before staining or added to ● hematoxylin dye to stain cell nuclei (and
dye solution other parts) blue
EXAMPLES: ● eosin dye to stain other structures pink or
● potassium alum with red
hematoxylin in Ehrlich's
hematoxylin ROUTINE H&E STAINING in Paraffin Embedded
● iron in Weigert's hematoxylin Section (Regressive Staining)
Fixation: Most fixatives can be used except osmic
● ACCENTUATOR - accelerates acid solutions which inhibit hematoxylin.
EXAMPLES: Procedure:
● potassium hydroxide in 1. Clear paraffin embedded sections in first
● Loeffler's methylene blue xylene bath for 3 minutes.
● phenol in carbol thionine and 2. Transfer to second xylene bath for 2 to 3
carbol fuchsin minutes.
3. Progressive ● tissue elements are stained in 3. Immerse in first bath of absolute ethyl
Staining a definite sequence, and the alcohol for 2 minutes.
staining solution is applied for

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HISTOPATHOLOGY - Histopathologic Techniques

4. Transfer to a bath of 95% ethyl alcohol for 1

or 2 minutes. XI. MOUNTING
5. Rinse in running water for 1 minute. ● MOUNTING MEDIUM: the solution in which
6. Stain with Harris alum hematoxylin for 5 the specimen is embedded, generally
minutes (Ehrlich's hematoxylin requires under a cover glass.
15-30 minutes). ● It may be liquid, gum or resinous, soluble in
7. Wash in running tap water to remove water, alcohol or other solvents and be
excess stain. sealed from the external atmosphere by
8. Differentiate in 1% acid-alcohol (1ml non-soluble ringing media
concentrated HCl to 99 ml. of 80% ethyl
alcohol) for 10-30 sec. monitoring the
changes in color microscopically until only
the nuclei are stained.
9. Rinse in tap water.
10. Blue in ammonia water (average of 5
minutes) or 1% aqueous lithium carbonate SPECIMENS FOR EXAMINATION
until the sections appear blue (about 30 I. Gynecological Performed regularly even
seconds). specimen in pregnant women
11. Wash in running water for 5 minutes. without undue risk
12. Counterstain with 5% aqueous eosin for 5 EXAMPLE: Vaginal smear
minutes. If alcoholic eosin is used, the time II. Non-gynecological EXAMPLE:
can be reduced to 30 seconds or 1 minute. specimen ● Respiratory Tract
13. If aqueous eosin is used, wash and specimens:
differentiate in tap water under microscope - Sputum
control until the nuclei appear sharp blue to - Bronchoalveolar
blue black and the rest of the tissue appear lavage (BAL)
in shades of pink. If alcoholic solution is III. Urine Determine the presence
used, differentiate with 70% alcohol. of urethral cancer
14. Dehydrate, clear and mount.
MOUTING STEPS:
H & E staining of Frozen Sections for Rapid 1. After cutting, sections are floated out on a
Diagnosis (Progressive Staining) water-bath.
1. Orient section in the block and freeze with a. Bubbles accumulating under the
liquid nitrogen. ribbon may be removed with a
2. Cut cryostat sections at 5-10 micron. smooth teasing needle, care being
3. Mount sections on to albuminized slides taken not to tear the section.
and dip in 10% formalin to fix. b. Bubbles may also be removed by
4. Rinse rapidly in water. pulling the ribbon very gently
5. Stain with Harris hematoxylin for 30-45 across the long edge of a glass
seconds. slide held below the section in the
6. Rinse in tap water. water bath.
7. Blue in ammonia water for 5 seconds. 2. When the sections chosen have flattened
8. Rinse in tap water. out, the numbered slide is immersed in the
9. Counterstain with 5% aqueous eosin or 1% water bath and the section is fished out
alcohol eosin for one minute. and drained.
10. Rinse in tap water. 3. After draining, the sections are fixed to the
11. Dehydrate in increasing concentrations of slides.
alcohol. a. This can be done either by leaving
12. Clear with xylene. the slides in a 37°C incubator
13. Mount with cover slide. overnight, by placing the slides in a

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HISTOPATHOLOGY - Histopathologic Techniques

wax oven at 56° to 60°C for 2 hours,

or by drying the slides on a hot
plate at 45° to 55°C for 30 to 45
minutes.
b. For more delicate tissues like the
CNS tissue or brain, a longer drying
time at lower temperature (e.g.
37°C for 24 hours or longer) is
recommended to avoid splitting
and cracking of the section due to
excess heat.
References
4. Another alternative to drying is by the use
● Histopathologic techniques ppt
of adhesives.
● Bruce-Gregorios, J.H. (1974)
Histopathologic Techniques. JMC Press
XII. LABELLING
Inc., Quezon City, Philippines, BAN CROFT,
● after sample collection, tissue is
Mahendra Jain A.C.P.M Dental College
accompanied by tag or label, bearing the
India.
lab. number given to specimen at start,
through all stages
○ Thin white card with writing in soft
pencil withstands all fluids used in
tissue processing.
● tissue specimen is received in the
laboratory have a request form that lists
the patient information, history &
description of the site of origin
○ the specimen are accessioned by
giving them a number that will
identify each specimen for each
patient

FERNING
● Arborization of mucus due to the formation
of salt crystals in high NaCl concentration
● Exhibits “palm leaf appearance”

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