Biopsy Type of Tissue

Obtained from LIVING source Tissue Structure to be Studied

Autopsy Type of stain to be used
obtained from DEAD source Effects of Fixative
FIXATION Harden tissue
Most Critical step Prevent Bacterial growth
Primary Goal of Fixation Reduce risk of infection
Preserve Morphology and Chemical Increase optical differentiation of cells and
Integrity of tissue as close to original tissue
Secondary Goal of Fixation Heat Fixation
To harden the tissue Thermal coagulation of Proteins
To protect the tissue from trauma of further usually for Bacterial smears and Frozen
handling sections
Fixation Time ACETONE
Minimum: 6 hrs Preserves Lipase, Phosphatase, Enzyme
Maximum: 48 hrs Studies
3 hrs ACETONE
Fixation Time for Electron Microscopy should be in ice cold temp. ( -5 -4 C)
Additive ACETONE and CARNOY'S FLUID
component becomes part of the tissue for diagnosis of RABIES
Non-Additive TRICHLOROACETIC ACID
Component does not become part but it Both Fixative and Decalcifying Agent
ALTERS the tissue component GLACIAL ACETIC ACID
20x the volume of the specimen (15-20:1) for nuclear proteins, nuclear chromatin.
amount of fixative cytoplasmic differentiation
5-10x GLACIAL ACETIC ACID
Amount of Fixative for Osmium Tetroxide SOLIDIFIES at 17 degrees celsius
20x ALCOHOL FIXATIVES
Amount of Fixative for Electron Microscopy both a Fixative and a Dehydrating Agent
NOT LESS THAN 50x ALCOHOL FIXATIVES
Amount of Fixative for Museum causes Glycogen Polarizing
Preparations Methyl Alcohol
Factors that will retard/prolonged Fixation for wet and dry prep. (blood smears and BM
Size and Thickness tissues)
Cold Temp. Ethyl Alcohol
Mucus and Blood (wash with NSS) for blood and tissue films
Fat (cut into thin slices) Isopropyl Alcohol
Factors that will Accelerate Fixation time for Touch Preparations
Size and Thickness Carnoy's Fluid
Agitation Most Rapid alcohol fixative
Heat and Pressure (37-56 C) Newcomer's
Factors to consider in choosing the compatible with Fuelgen
appropriate fixative Newcomer's - for mucopolysaccharides and
Need for immediate examination nuclear CHON

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10% Formaldehyde PICRIC ACID
used for mailing specimens Both FIXATIVE and STAIN
10% Formaldehyde Bouin's Fluid
precipitation of White Paraformaldehyde is Abolishes Fuelgen reaction
possible Bouin's Fluid
10% Formaldehyde for Pituitary biopsies, Embryo, Endometrial
aldehyde fixative which may cause curettings
dermatitis Bouin's Fluid
10% Formaldehyde not suitable for kidney
has a 1mm/hr penetration rate Brasil's Alcoholic Picroformol
37 - 40% Formaldehye Excellent fixative for GLYCOGEN
"100% formaldehyde" LEAD Fixatives
Gendre's Fluid for ACID MUCOPOLYSACCHARIDES and
aldehyde fixative which fixes sputum CT mucin
Gendre's Fluid Chromic acid
used for microincineration technique fixative for carbohydrates
Formol Corrosive 3% potassium dichromate
100% formalin + mercuric chloride preserves lipids and mitochondria
Formol Corrosive Regaud's/Moller's
recommended for Neutral fats, for Chromatin, Golgi Bodies, Mitochondria,
Phospholipids, lipids and RBC containing organs
10% Formol Saline Orth's fluid
aldehyde fixative used for CNS, fats and for early degenerative processes and tissue
enzymes necrosis
10% Formol Saline Orth's Fluid
aldehyde fixative which is for post-mortem fixes Rickettsia org.
tissue MERCURIC Fixatives
Glutaraldehyde excellent for TISSUE PHOTOGRAPHY and
aldehyde fixative for electron microscopy TRICHROME staining
and enzyme histochemistry Zenker's Fluid
2.5% Glutaraldehyde for liver, spleen, CT, fibers, nuclei
glutaraldehyde for smaller specimens Zenker's Formol/HELLEY'S
4% Glutaraldehyde for PITUITARY gland, BM and other blood
glutaraldehyde for big specimens containing organs
Karnovsky's Heidenhan's Susa
paraformaldehyde-glutaraldehyde and For TUMOR biopsies
ACROLEIN B5 Fixative
aldehyde fixative for electron microscopy for Bone Marrow
and immunochemistry B5 Fixative
PICRIC ACID contains Anhydrous Sodium Acetate
imparts yellow discoloration Flemming's
Acid Lithium Carbonate osmium tetraoxide fixative that contains
treatment to remove yellow color in picric acetic acid
acid fixatives

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Flemming's methyl alcohol
excellent for NUCLEAR structure dehydrating agent used for blood and tissue
Flemming's without acetic acid smear preparations
for CYTOPLASMIC structure butyl alcohol
Microwave Technique for plants and animals
accelerates FIXATION, isopropyl alcohol
DECALCIFICATION, STAINING dehydrating agent substitute for ethanol or
Microwave Technique xylene
for neurochemical substances in the brain isopropyl alcohol
like acetylcholine used in microwave technique
SECONDARY FIXATION Acetone
purposes: rapid acting dehydrating agent for small
make special staining possible pieces of tissue
complete hardening and preservation of Dioxane
tissue excellent dehydrating and clearing agent
improve demonstration of desired subs Graupner's
Post-Chromatization 3 changes of pure dioxane and 3 changes
a secondary fixation that uses any chromate of paraffin
fixative Weiseberger's
WASHING OUT like graupner's but is wrapped with gauze
removes excess fixatives bag
tap water calcium oxide
removes excess osmic acid and chromates absorbs water in the weiseberger's method
50-70% alcohol ethylene glycol/ cellosolve
removes excess picric acid Abortion (toxic to fetal and reproductive
Alcoholic Iodine system)
removes excess mercuric fixative ethylene glycol/cellosolve
DEHYDRATION combustible at 100-120 F
not less than 10x the volume of the tetrahydrofuran
specimen for atheromatic arteries and heart
DEHYDRATION collagenous
70% alcohol to ascending grades of alcohol tetrahydrofuran and carboxyl
dehydration both a dehydrating and clearing agent
ascending grades of OH CLEARING
hydration removes alcohol and make tissue
descending grades of alcohol transparent
Anhydrous Copper Sulfate clearing agent must miscible with:
indicator that will accelerate the process Dehydrating agent
Blue color Infiltrating agent
color when it is fully/completely dehydrated Mounting medium
Ethyl alchohol factors affecting clearing
routine dehydrating agent viscosity
ethyl alcohol boiling point
best dehydrating agent prolonged exposure to clearing agent

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gum syrup and glycerine carbon disulfide and Phenol
clearing agent for frozen sections clearing agent for smooth muscles
xylene carbon xylene
miscible with alcohol and paraffin clearing agent for friable tissue
xylene creosote
carcinogenic reagent clearing agent for celloidin sections
xylene high test aviation lead free gasoline
most rapid clearing agent for urgent biosies also an excellent clearing agent
toluene/toluol DECALCIFICATION
subsitute for xylene or benzene calcium ion or lime salts are removed from
toluene tissues
tends to acidify in a partially filled vessel 1-3 mm
benzene ideal thickness for decalcification
best clearing reagent for embedding decalcification
process prevent obscuring the microanatomical
benzene detail
cause damage to BM and kidney Nitric acid
benzene most common decalcifying agent
cause aplastic anemia 10% aqueous nitric acid
chloroform recommended for heavy cortical bones
only clearing agent that does not make 10% aqueous nitric acid and formol nitric
tissue transparent acid
chloroform decalcifying agent that imparts a yellow
toxic to the liver color
cedarwood oil nitrous acid
clear both paraffin and celloidin sections treatment to remove yellow color for 10%
cedarwood oil aqueous nitric acid
clearing agent recommended for cytological 0.1% urea
studies treatment to remove yellow color for formol
clove oil nitric acid
tends to become adulterated 5% sodium sulfate
aniline oil neutralizes the tissue when using formol
for embryos, insects and delicate tissues nitric acid
methyl benzoate perenyi's fluid
used when double embedding technique is nitric acid + ethanol + chromic acid (NED)
required perenyi's fluid
bergamot decalcifies and softens tissue
clearing agent for skin and smooth muscles perenyi's fluid
origanum slow decalcifying agent for dense bones
clearing agent for skin phloroglucinol nitric acid
wintergreen most rapid decalcifying agent
clearing agent for delicate tissue hydrochloric acid
terpineol decalcifying agent used for surface
clearing agent for eyes decalcification of blocks

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von ebner's fluid most sensitive and reliable method to
decalcifying agent that permits relatively measure extent of decal
good cytological staining opacity
von ebner's fluid indicator of incomplete decal. in radiologic
for teeth and small.pieces of bones method
formic acid chemical method
used for routine decalcification of post use ammonium oxalate to measure the
mortem research studies extent of decal
citrate turbidity
component added to chelates calcium in indicator of incomplete decal. in chemical
decalcification using formic acid method
chromic acid lithium carbonate
both fixative and decalcifying agent used to wash tissue.in post decalcification
chromic acid 30 mins
for minute bone spicules time to remove acids after decal for small
chromic acid tissues
environmental toxin 1-4 hours
chloroform time to remove acids after decal. for large
preservative for citric acid citrate buffer specimens
EDTA factors influencing rate of decal
chelating agent excellent for electron concentration.and volume
microscopy heat
EDTA structure and tep
inactivates ALP mechanical agitation
Magnesium Chloride tissue softeners
substance added in EDTA to prevent molliflex
inactivation of ALP perenyi's
1-3 weeks lendrums method
decal. time for small specimens using EDTA mollifex
6-8 weeks 1% HCl in 70% alcohol
decal. time of dense tissues using EDTA perenyi
ion exchange resin 2% HCl in 70% alcohol
remove calcium from formic acid containing lendrums method
decal agents 4% aqueos phenol
ammonium polystyrene IMPREGNATION
resin used to remove calcium in ion clearing agent is removed
exchange resin impregnation
electrophoresis provides a firm consistency to the specimen
most rapid method of decal. for easier handling and cutting
physical method xylene and benzene
inaccurate method that measures extent of clearing agent easily removed
decal. that damages tissue chloroform and cedarwood oil
radiologic method clearing agent difficult to remove

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paraffin wax embedding eyes
most common used for impregnation tissue mat
paraffin wax paraffin with rubber
not recommended for fatty tissues carbowax
paraffin wax water soluble wax
allows cutting of serial sections carbowax
56 degrees celsius wax recommended for neuropathological
melting point for routine work for tissues
impregnation carbowax
54-58 degrees celsius wax that eliminates dehydration and
melting point if laboratory temp is between clearing
20-24 degrees celsius celloidin
50-54 degrees celsius purified form of nitrocellulose
melting point if laboratory temp is between celloidin impregnation
15-18 degrees celsius impregnating medium for tissues with large
manual method hollow cavities that tend to collapse
method of impregnation that involves 4 celloidin
changes of paraffin medium where serial sections are difficult to
2-5 degrees celsius higher than the melting cut
point or 56-60 degrees celsius celloidin
temp range for paraffin oven in medium where photomicrograph is difficult
impregnation to obtain
automic method wet celloidin
method that involves 2-3 changes of celloidin that uses 70% alcohol and is
paraffin recommended for bones and brains
vacuum dry celloidin
infiltration under negative atmospheric celloidin method for eye specimens
pressure gilson's mixture
paraplast chloroform + cedarwood oil
a mixture of highly purified paraffin and gelatin
synthetic polymer impregnating medium for histochemical and
56-57 degrees celsius enzyme studies
melting pt of paraplast 1% phenol
56-58 degrees celsius added to gelatin to prevent molds
melting pt of embedded plastic resin
embeddol impregnating medium for hard tissues like
similar to paraplast but less brittle and less uncalcified bones
compressible light microscope
46-48 degrees celsius microscope used to provide superior results
melting pt of ester wax using plastic resin
ester wax polyester
harder than paraffin and uses heavy duty plastic resin for electron microscopy
microtome acrylic
bioloid plastic resin that uses light microscopy

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leuckhart's -20 degrees celsius
2 L shaped strip of heavy brass metal average temp of the cryostat
compound embedding 10-12 micrometers
series of interlocking spaces used for size of tissue using rocking microtome
embedding 4-6 micrometers
5-10 degrees celsius higher than the size of tissue using rotary microtome
melting point 7-9 micrometers
temperature of melted paraffin for size of tissue using the sliding microtome
embedding 10-15 micrometers
double embedding size of tissue using freezing microtome
infiltrate with celloidin and embedded with 4 micrometers
paraffin size of tissue using cryostat
block holder 0.5-1 micrometer
part of the microtome where the tissue is tissue size using ultrathin microtome
hold in position plane concave
paldwell one flat size one concave that is 25 mm
invented rocking microtome long
minot plane wedge
invented rotary microtome both sides are flat and is 100 mm long
adams biconcave
developed sliding microtome both sides are concave that is 120 mm long
quekett bevel angle
invented freezing microtome formed between cutting edges (27-32)
standard sliding microtome clearance angle
most dangerous microtome formed between surface of block and cutting
rocking microtome edge of knife (0-15)
most simplest microtome wedge angle
rotary microtome formed by the sides of wedge knife(14-15)
most common microtome honing
knife grinding the cutting edge to acquire even
movable part in standard sliding microtome size edge
freezing microtome heel to toe
uses intermittent burst of CO2 movement of honing
cryostat or cold microtome remove nicks
for fresh tissue microtome purpose of honing
cryostat 20-30
microtome for fluorescent antibody staining strokes for honing
and histochemical enzyme studies belgium yellow
rotary microtome gives best results of honing
microtome inside a cryostat arkansas
ultrathin microtome hone that shows more polishing effects
microtome for electron microscopy fine carborundum
diamond knife for badly nicked knives
knife for ultrathin microtome

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lubricants for honing mountant that does not allow to view the
soapy water slide under OIO
mineral oil kaiser
clove oil other name for glycerin jelly
xylene glycerin jelly or kaisser's
stropping requires ringing
polishing and sharpening the cutting edge apathy's medium
stropping mountant for methylene blue stained nerve
remove burrs preparation
toe to heel brun's fluid
movement of stropping mountant for frozen sections
40-120 strokes 1.46
number of strokes for.honing glycerin refractive index
horse leather 1.47
material used to make a paddle strop glycerin jelly refractive index
vegetable oil or castor oil 1.43
treatment used for the paddle strop farrant's medium refractive index
mineral oil 1.52
oil which should not be used to treat the apathy's medium refractive index
paddle strop resinous mounting medium
fishing out mountant for tissue that have been
removal of tissue ribbon from the float out dehydrated and cleared
bath Abus balsamia
5-10 degrees celsius lower than melting tree where Canada balsam was extracted
point DPX
temperature of the floating water bath mountant only for small tissue sections
mayer's egg albumin 1.524
egg white + glycerine canada balsam refractive index
mayer's egg albumin 1.532
most common adhesive DPX refractive index
thymol XAM
subs added to adhesives to prevent growth 1.52
of molds Clarite
APES 1.544
adhesive recommended for cytology ringing
mounting sealing the margins of the coverslip to
process of adding syrupy fluid on the slide prevent escape of fluid or semi fluid mounts
1.158 Durofix
refractive index of theglass slide ringing medium for cellulose adhesive
aqueous mounting medium kronig cement
temporary mountant 2 parts paraffin wax + 4-9 parts of
resinous mounting media colophony resin
permanent mountant deparaffinization
water

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removal of paraffin wax once fixed on the Phenol
slide accentuator of carbol fuchsin and carbol
Staining thionin
process to study the architectural pattern of progressive staining
the tissue No differentiation and decolorization is
histological staining involved
tissue has a direct interaction with the dye progressive staining
histological staining gradual application if the dye
staining that produces coloration of the regressive staining
active tissue component tissue is overstained first
histochemical staining decolorization
tissue is demonstrated thru chemical process where excess stain is removed in
reactions regressive staining
Perl's prussian blue metachromatic staining
stain for hemosiderin staining tissue with a color that is different
Periodic acid schiff from the stain itself
stain for carbohydrates counterstaining
active agent method of staining that produce contrast
it is the substrate in the enzyme and background
histochemistry of staining metallic impregnation
immunohistochemical staining tissue elements are demonstrated by
stain that detects phenotypic markers colorless solutions of metallic salts
antibodies black deposits
it is detected in the immunohistochemical substance that may tend to be produced in
staining for detection of phenotypic markers metallic impregnation
direct staining vital staining
staining method that uses aqueous or staining for live cell constituents
alcoholic dyes nucleus
indirect staining resistant using the vital staining
staining method that is intensified by adding intravital staining
another agent stain is injected to any part of the living body
mordant supravital staining
substance that serves as a link or bridge stain is applied immediately after removal of
between the tissue and the dye cells from the living body
accentuator ammonia water
accelerates or hastens the staining process bluing agent
potassium alum nucleus
mordant of erlich's hematoxylin part of cell that is stained by hematoxylin in
ferric ammonium chloride H&E staining
mordant of weigert's hematoxylin ammonia water
ferric ammonium sulfate intensifies the color of the nucleus in H&E
mordant of heidenhain's hematoxylin staining
KOH cytoplasm
accentuator of loeffler's methylene blue part of cell that is stained by eosin in H&E

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blue to blue black cochineal dyes
color of nucleus in H&E dyes extracted from bugs treated with alum
dark blue carmine
color of karyosome in H&E product when bugs for cochineal dyes are
purplish blue treated with alum
color of calcium and decalcified bones in picro carmine
H&E dye recommended for neuropathy studies
bright orange best carmine
color of RBC, keratin and eosinophilic dye that contains aluminum chloride and
granules in H&E used for elastic fibers
pale pink synthetic dyes
color of cytoplasm in H&E "coal tar dyes"synthetic dyes
pink or deep pink dyes from hydrocarbon benzene
color of muscle fibers in H&E chromophore
pink responsible for coloring property
color of decalcified bone matrix, collagen, auxochrome
and osteoid in H&E responsible for dying property
natural dye amphoteric dyes
stain that is derived from plants and animals dyes with a neutral pH
hematoxylin lysochrome dyes
formed by ripening or from adding oxidizing oil soluble dyes
agents sudan black
hematein most sensitive, dye with a greater affinity to
active coloring agent of hematoxylin phospholipids and neutral fats
alum hematoxylin sudan III
use potassium alum as a mordant first sudan dye, for central nervous system
sodium iodate tissues
ripening agent of erlich's and mayer's sudan IV
hematoxylin dye for trigylcerides
mercuric oxide benzoic acid
ripening agent of harris' hematoxylin intensifies fat and prevents rapid
alcoholic iodine deterioration of the solution
ripening agent of cole's hematoxylin
iron hematoxylin
usually used for photomicrography
weigert's hematoxylin
hematoxylin for muscles and CT fibers
heidenhain's hematoxylin
hematoxylin for nuclei and cytoplasmic
inclusions
copper hematoxylin
for spermatogenesis studies
tungsten hematoxylin/PTAH
use 1% phosphotungstic acid as a mordant

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