Chapter 3: FIXATION AND FIXATIVES | 1
I. ALDEHYDE FIXATIVES
FIXATIVE DESCRIPTION
 10% conc. is widely used
 made from the oxidation of
methyl alcohol
 37 – 40% w/v soluble in
water
 35%-40% gas by weight
 buffered to pH 7
(phosphate buffer)
Fixation time: 24 HOURS
Formaldehyde
(Formalin)
 Precipitates may be
removed by filtration or by
addition of 10% methanol
 Bleaching of tissues may be
prevented by changing the
fluid fixative every three
months
PURPOSE
 preserves fat and mucin
 preserves glycogen
 preserves proteins
 recommended for nervous tissue
preparation
ADVANTAGES
 cheap, readily available, easy to prepare,
relatively stable
 compatible with many stains
 does not overharden tissues
 penetrates tissues well
 does not precipitate proteins
 does not make tissues brittle
 colored tissue photography
 allows frozen tissue sections to be prepared
easily
 DOESN’T REQUIRE WASHING OUT
 Used for mailing specimens
DISADVANTAGES
 Fumes are irritating to nose and eyes
 Solution is irritating to skin (causes allergic
dermatitis on prolonged contact)
 May produce considerable shrinking of
tissues
 Does not harden some cytoplasmic
structures adequately
 If unbuffered:
 formalin reduces both basophilic and
eosinophilic staining of cells (acidity of
formic acid may be used when applying
the silver impregnation technique of
staining)
 forms abundant brown pigment granules
on blood containing tissues due to
blackening of hemoglobin
 Prolonged fixation:
 bleaching of specimen/loss of natural
colors
 dispersing of fat from the tissues into the
fluid
 dissolution or loss of glycogen, biurate of
sodium crystals and uric acid
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 Simple microanatomical
fixative
 Made up of saturated
formaldehyde (40% w/v)
Fixation time:
24 HOURS at 35˚C (95˚F)
10% Formol-Saline 48 HOURS at 20-25˚C (65-77˚F)
 most common fixative
Formula:
3.5mg
Sodium dihydrogen
phosphate (anhydrous)
10% Neutral 6.5mg
Buffered Formalin/ Disodium hydrogen
Phosphate- phosphate (anhydrous)
Buffered Formalin 100ml
40% Formaldehyde
900ml
Distilled Water
Fixation Time: 4 – 24 HOURS
Fixation time: 3 – 24 HOURS
Formal-Corrosive
(Formal Sublimate)
 Recommended for fixation of central
nervous tissues & general post-
mortem tissues for histochemical
examinations
 recommended for preservation and
storage of surgical, post-mortem and
research specimen
 formol-mercuric chloride solution is
recommended for routine post-
mortem tissues
 penetrates and fixes tissues evenly
 preserves microanatomic and cytologic details
with minimum shrinkage and distortion
 large specimens may be fixed for a long time
provided that the solution is changed every
three months
 preserves enzymes and nucleoproteins
 demonstrates fats and mucin
 does not overharden tissues; facilitates
dissection of specimen
 ideal for most staining techniques (including
SILVER IMPREGNATION)
 allows natural color of tissue to be restored
upon immersion in 70% alcohol
 prevents precipitation of acid formalin
pigments on post-mortem tissue
 best fixative for tissues containing iron
pigments and for elastic fibers which do not
stain well after Susa, Zenker or Chromate
fixation
 requires no post-treatment after fixation, so
the tissue goes directly to 80% alcohol for
processing
 penetrates small pieces of tissue rapidly
 produces minimum shrinkage and hardening
 excellent for many staining procedures
including SILVER RETICULUM METHODS
 brighten cytoplasmic and metachromatic
stains better than with formalin alone
 cytologic structures and blood cells are well
preserved
 no need for “washing out”; tissues can be
transferred directly from fixative to alcohol
Chapter 3: FIXATION AND FIXATIVES | 2
Similar to formalin with the following additions:
 slow fixative (≥ 24 hours)
 tissues tend to shrink during alcohol
dehydration (this may be reduced by
secondary fixation)
 Metachromatic reaction of amyloid is
reduced
 Acid dye stains less brightly than when fixed
with mercuric chloride
 longer to prepare; time-consuming
 positivity of mucin to Periodic acid-Schiff
(PAS) stain is reduced
 may produce gradual loss in basophilic
staining of cells
 reactivity of myelin to Weigert’s iron
hematoxylin stain is reduced
 INERT TOWARDS LIPIDS, especially neutral
fats and phospholipids
 penetration is slow (tissue sections should
not be more than 1 cm thick)
 forms mercuric chloride deposits
 does not allow frozen tissue sections to be
made
 inhibits the determination of the extent of
tissue decalcification
@ohkeye & 楊桂霞

 post-fixation with phenol-
formalin for 6 hours or
more can enhance
immunoperoxidase studies
on the tissues and in some
Alcoholic Formalin cases, electron microscopy,
(Gendre’s) Fixative if it is necessary at a later
time to establish a diagnosis
 made up of two
formaldehyde residues,
linked by three carbon
chains
 acts similarly to
formaldehyde
 sometimes used for routine
light microscopy
Glutaraldehyde  buffered glutaraldehyde,
followed by secondary
fixation in osmium
tetroxide is satisfactory for
electron microscopy
Fixation time: varies from ½
hour (for small specimens) to
2 hours (maximum
contribution)
II. METALLIC fixatives
A. Mercuric chloride
FIXATIVE DESCRIPTION
 most common metallic
Mercuric Chloride fixative
 used to fix sputum since it
coagulates mucus
 2.5% solution – small tissue
fragments and needle biopsies
Fixation time: 2 – 4 HOURS
 4% solution – large tissues less than
4mm thick
Fixation time: 6-8 HRS up to 24 HRS
PURPOSE
 recommended for renal tissues,
fibrin, connective tissues & muscles
 fixes lipids; especially neutral fats and
phospholipids
 fixation is faster (reduced to one-half)
 can be used for rapid diagnosis since it fixes
and dehydrates at the same time (i.e. frozen
section)
 good from preserving glycogen and for micro-
incineration technique
 more stable effects on tissues, giving a firmer
texture with better tissue sections; especially
of nervous tissues
 preserves plasma proteins better
 produces less tissue shrinkage
 preserves cellular structures
 recommended for enzyme histochemistry and
electron microscopy
 more pleasant and less irritating to the nose
 does not cause dermatitis
ADVANTAGES
 penetrates & hardens tissues rapidly and well
 nuclear components are shown in fine detail
Chapter 3: FIXATION AND FIXATIVES | 3
 produces gross hardening of tissues
 causes partial lysis of RBCs
 preservation of iron-containing pigments is
poor
 formaldehyde foes not give as good a
morphological picture as glutaraldehyde
 it causes little cross-linking under usual
fixation conditions where low concentration
of proteins are used (glutaraldehyde is
more effective at cross-linking)
 more expensive
 less stable
 penetrates tissues more slowly
 tends to make tissue (i.e. renal biopsy) more
brittle
 reduces PAS positivity of reactive mucin
(may be prevented by immersing
glutaraldehyde-fixed tissues in a mixture of
concentrated glacial acetic acid and aniline
oil)
PRECAUTIONS:
 specimen vial must be kept refrigerated
during fixation process
 swirling the vials ensures that the
specimen is in contact with the fresh
solution at all times as it might change
DISADVANTAGES
 penetrates poorly & causes marked shrinkage
of cells, so it’s usually combined with other
@ohkeye & 楊桂霞

 frequently used in sat. aq.
solutions of 5-7%
 widely used as a secondary
fixative, reacting with a
number of amino acid
residues & accompanied by
spectroscopic changes,
probably due to reactions
with histidine residues
 compound solutions must
always be freshly prepared
 use of metallic forceps and
metallic caps should be
avoided
 contact of fixative with
personal jewelries should
be avoided
 made up of mercuric
chloride stock solution to
which glacial acetic acid is
added to it just before use
to prevent turbidity &
formation of dark
precipitate
Zenker’s Fluid  solutions must always be
freshly prepared
 tissues should be cut thin
(2-3mm) & hollow organs
should be opened to
promote complete
penetration & fixation
 good general preservative for all
kinds of tissues
 gives excellent staining results
 recommended for fixing small pieces
of liver, spleen, connective tissue
fibers & nuclei
 precipitates all proteins
 great affinity to acid dyes and is preferred in
lieu of formaldehyde for cytoplasmic staining
 Trichome staining is excellent
 routine fixative of choice for preservation of
cell detail in tissue photography
 permits brilliant metachromatic staining of
cells
 produces fairly rapid and even fixation
 stock solutions keep well without
disintegration
 recommended for Trichrome staining
 permits brilliant staining for nuclear &
connective tissue fibers
 compatible with most stains
 may act as a MORDANT to make certain
special staining reactions possible
Chapter 3: FIXATION AND FIXATIVES | 4
fixative agents (this may be counteracted by
addition of acid)
 rapidly hardens the outer layer of the tissue
with incomplete fixation of the center (thin
sections should be made)
 penetration beyond the first 2-3mm is slow
(not more than 5mm thickness of tissues
should be used)
 if left in the fixative for more than 1-2 days,
the tissue becomes unduly hard and brittle
 prevents adequate freezing of fatty tissues
and makes cutting of frozen tissue difficult
 causes considerable lysis of RBCs and
removes iron from hemosiderin
 INERT TO FATS AND LIPIDS
 leads to the formation of black granular
deposits in the tissues (may be removed by
adding sat. iodine solution in 90% alcohol;
iodine will be decolorized with absolute
alcohol in the subsequent stages of
dehydration)
 reduces the amount of demonstrable
glycogen
 compound solutions deteriorate rapidly upon
addition of glacial acetic acid to formalin
 extremely corrosive to metals
 penetration is poor
 not stable after addition of acetic acid
 prolonged fixation (for more than 24 hours
(will make tissues brittle and hard
 causes lysis of RBCs & removes iron
hemosiderin
 does not permit cutting of frozen sections
 tendency to form mercuric pigment deposits
or precipitates (may be removed by de-
zenkerization)
 Bring slides to water
 Immerse in Lugol’s iodine (5 min.)
 Wash in running water (5 min.)
 Immerse in sodium thiosulfate 5% (5 min.)
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Fixation time: 12 – 24 HOURS
Fixation time: 12 – 24 HOURS
Zenker-formol
(Helly’s solution)
 excellent cytologic fixative
 after using this fixative, the
tissue should be transferred
directly to a high-grade
alcohol (i.e. 96% or
absolute alcohol) to avoid
Heidenhain’s Susa undue swelling of tissues
Solution caused by treatment with
low-grade alcohol
Fixation time: 3 – 12 HOURS
 prior to use, add 1cc. of
formaldehyde (40%) for
10cc. of B-5
B-5 Fixative
Fixation time: 1 ½ HR – 2 HRS
B. CHROMATE FIXATIVES
FIXATIVE DESCRIPTION
 excellent microanatomic fixative for
PITUITARY GLAND, bone marrow,
spleen & liver
 recommended mainly for TUMOR
biopsies (especially of the skin)
 used for BONE MARROW BIOPSIES
PURPOSE
 penetrates and fixes tissues well
 nuclear fixation & staining is better than
Zenker’s
 preserve cytoplasmic granules well
 penetrates and fixes tissues rapidly & evenly
 produces minimum shrinkage and hardening
of tissues due to the counter-balance of the
swelling effects of acids and the shrinkage
effect of mercury
 permits most staining procedures to be done,
including silver impregnation, producing
brilliant results with sharp nuclear &
cytoplasmic details
 permits easier sectioning of large blocks of
fibrous connective tissues
 Susa-fixed tissues may be transferred directly
to 95% alcohol or absolute alcohol, thereby
reducing processing time
 good fixative for cytology of bone marrow
biopsies
ADVANTAGES
Chapter 3: FIXATION AND FIXATIVES | 5
 Wash in running water (5 min.)
 Proceed with required water-soluble stain
 tissue must be washed in running water for
several hours (or overnight) before
processing
 insufficient washing may inhibit or interfere
with good cellular staining
 similar to Zenker’s
 brown pigments are produced if tissues
(especially blood containing organs) are
allowed to stay in the fixative for more than
24 hours due to RBC lysis (may be removed
by immersing the tissue in sat. alcoholic
picric acid or sodium hydroxide)
 prolonged fixation of thick materials may
produce considerable shrinkage, hardening
and bleeding (tissues should not be >1mm)
 RBC preservation is poor
 some cytoplasmic granules are dissolved
 mercuric chloride tend to form in tissues
(these may be removed by immersion of
tissues in alcoholic iodine solution)
 Weigert’s method of staining elastic fibers is
not possible in Susa-fixed tissues
 overfixation hardens the tissue and makes
cutting difficult
 some B-5 solutions will form precipitate on
standing (but this is of no consequence)
 may causing mercuric pigments to form on
tissues (de-zenkerization may be employed)
DISADVANTAGES
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 used in 1-2% aq. solution,
usually as a constituent of a
compound fixative
Chromic Acid
 used in a 3% aq. solution
Potassium
Dichromate
Fixation time: 12 – 48
Redard’s (Muller’s
Fluid)
• strong formaldehyde (40%)
is added just before use
Orth’s Fluid
Fixation time: 36 – 72 HOURS
C. LEAD FIXATIVES
FIXATIVE DESCRIPTION
 used in 4% aq. solution of
Lead Fixatives
basic lead acetate
 recommended for demonstration of
chromatin, mitochondria, mitotic
figures, Golgi bodies, RBC & colloid-
containing tissues
 demonstrates RICKETTSIAE & other
bacteria
PURPOSE
 precipitates all proteins
 adequately preserve carbohydrates
 fixes but does not precipitate cytoplasmic
structures
 preserves lipids
 preserves mitochondria (if used in pH 4.5 –
5.2)
 penetrates tissues well
 hardens tissues better and more rapidly than
Orth’s fluid
 recommended for the study of degenerative
processes & tissue necrosis
 preserves myelin better than buffered
formalin
ADVANTAGES
 recommended for acid mucopolysaccharides
 fixes connective tissue mucin
Chapter 3: FIXATION AND FIXATIVES | 6
 it’s a strong oxidizing agent (a strong
reducing agent, such as formaldehyde, must
be added to chrome-staining fixatives before
use in order to prevent counteracting effects
& consequent decomposition of solution
upon prolonged standing)
 if solution becomes acidified, cytoplasm,
chromatin bodies & chromosomes are fixed
but mitochondria is destroyed
 deteriorates & darkens on standing due to
acidity (solution must always be freshly
prepared)
 penetration is slow (tissues should be no
thicker than 2 – 3 mm)
 chromate fixed tissues tend to produce
precipitates od sub-oxide (tissues should be
thoroughly washed in running water prior to
dehydration)
 prolonged fixation blackens tissue pigments
like melanin (tissues should be washed in
running water prior to dehydration)
 glycogen penetration is poor; generally
contraindicated for carbohydrates
 nuclear staining is poor
 does not preserve fats
 preserved hemosiderin less than buffered
formalin
 intensity of PAS reaction is reduced
 same as Regaud’s fluid
DISADVANTAGES
 takes up CO2 to from insoluble lead carbonate
especially upon prolonged standing (removed
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 Chapter 3: FIXATION AND FIXATIVES | 7
by filtration or adding acetic acid drop by drop
to lower the pH and dissolve the residue)

III. PICRIC ACID fixatives

FIXATIVE DESCRIPTION PURPOSE ADVANTAGES DISADVANTAGES
 Normally used in sat. aq.  excellent fixative for glycogen  stable, penetrates tissues well & fixes small  dyes tissues yellow (dye may be removed by
solution (approx.. 1%) demonstration tissues rapidly treatment with another acid dye or lithium
 yellow dye may be removed  yellow stain prevents small fragments from carbonate)
by treatment with another being overlooked  excessive staining may be removed by
acid dye or lithium  allows brilliant staining with the trichrome placing the tissues in 70% ethanol followed
carbonate method by 5% sodium thiosulfate & washed with
 suitable for Aniline stains (Mallory’s, running water
Heidenhain’s or Masson’s methods)  causes RBC hemolysis & reduces amount of
 precipitates all proteins demonstrable ferric iron in tissues
 not suitable for frozen sections because they
will crumble when cut
 prolonged fixation makes tissues hard, brittle
& difficult to section (not longer than 12 – 24
Picric Acid
hours; depending on size)
 picrates are formed upon protein & are soluble
in water (tissues must be rendered insoluble
by direct immersion in 70% alcohol)
 fixed tissues must never be washed in water
prior to dehydration
 picric acid is highly explosive when dry (keep
moist with distilled water or sat. alcohol at
0.5 to 1% conc. during storage)
 alters and dissolves lipids
 interferes with Azure eosin method of staining
(tissues should be thoroughly washed with
alcohol)

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 fc
Fixation time: 6 – 24 HOURS
Bouin’s Solution
Brasil’s Alcoholic
Picroformol
Fixative
GLACIAL ACETIC ACID
FIXATIVE DESCRIPTION
 colorless liquid
 called “Glacial” Acetic Acid
when undiluted because it is
a water-free (anhydrous)
acetic acid that freezes and
Acetic Acid solidifies at about 17°C.
 recommended for fixation of
embryos & pituitary biopsies
PURPOSE
 Normally used in conjunction with
other fixatives to form a compound
solution
 It precipitates DNA, which is split off
from nucleoprotein
 valuable for the preservation of
nuclei
Chapter 3: FIXATION AND FIXATIVES | 8
 produces minimal distortion of  penetrates large tissues poorly
microanatomical structures  picrates are soluble in water (tissues must be
 can be used for general & specific stains transferred directly from fixative to 70%
(shrinking effect of picric acid is balanced by alcohol)
the swelling effect of glacial acetic acid)  not suitable for fixing kidney structures, lipids
 excellent fixative for preserving soft & & mucus
delicate structures (i.e. endometrial  destroys cytoplasmic structures (i.e.
curettings) mitochondria)
 penetrates rapidly & evenly, causing little  produces RBC hemolysis & removes
shrinkage demonstrable ferric iron from blood pigments
 yellow stain is useful when handling  reduces or abolishes Feulgen reaction due to
fragmented biopsies hydrolysis of nucleoproteins
 permits brilliant staining of tissues
 preferred fixative for tissues to be stained by
Masson’s trichrome for collagen, elastic or
connective tissue (if tissue is fixed in
formalin, a pre-treatment of Bouin, as a
mordant prior to trichrome stain, is
recommended)
 preserves glycogen
 does not need “washing out”
 better and less messy than Bouin’s solution
 excellent fixative for glycogen
ADVANTAGES DISADVANTAGES
 It fixes and precipitates nucleoproteins.  When combined with Potassium Dichromate,
 It precipitates chromosomes and chromatin the lipid-fixing property of the latter is
materials; hence, is very useful in the study of destroyed (e.g. Zenker's fluid).
nuclear components of the cell. In fact, it is an  It is contraindicated for cytoplasmic fixation
essential constituent of most compound since it destroys mitochondria and Golgi
nuclear fixatives. elements of cells.
 It causes tissues (especially those containing
collagen) to swell. This property is used in
certain compound fixatives to counteract the
shrinkage produced by other components
(e.g. mercury).
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 Chapter 3: FIXATION AND FIXATIVES | 9
Alcohol fixatives
FIXATIVE DESCRIPTION PURPOSE ADVANTAGES DISADVANTAGES
 It is excellent for fixing dry and wet smears,  Penetration is slow.
Methyl Alcohol blood smears and bone marrow tissues.  If left in fixative for more than 48 hours,
100%  It fixes and dehydrates at the same time. tissues may be over hardened and difficult to
cut.
 is used for fixing touch preparations
 some touch preparations are air
Isopropyl Alcohol
dried and not fixed, for certain
95%
special staining procedures such
as Wright-Giemsa
 is used at concentrations of  It may be used as a simple fixative.  Preserves but does not fix glycogen.  Causes polarization of glycogen granules
70-100%  frequently incorporated into  Fixes blood, tissue films and smears.  Produces considerable hardening and
 use of lower concentrations compound fixatives for better results  Preserves nucleoproteins and nucleic acids; shrinkage of tissues
cause hemolysis of RBC and used for histo-chemistry, especially for  Hemosiderin preservation is less than in
Ethyl Alcohol inadequately preserved WBC enzyme studies. buffered formaldehyde
 Fixes tissue pigments fairly well.  A strong reducing agent; should not be mixed
Fixation Time: 18-24 HOURS w/ strong oxidizing agents such as chromic
acid, potassium dichromate& osmium
tetroxide
 Rapid in action  Recommended for fixing  Most rapid fixative; may be used for urgent  Produces RBC hemolysis
 Tissues fized in Carnoy’s for 1 chromosomes, lymph glands & urgent biopsy specimens for paraffin processing w/in  Causes considerable tissue shrinkage
hr can be transferred directly biopsies 5 hrs  Suitable ONLY for small pieces of tissues due
to absolute alcohol, or an  May be used for urgent biopsy spx for  Fixes and dehydrates at the same time. to slow penetration
alcohol-chloroform mixture paraffin processing w/in 5 hrs  Permits good nuclear staining and  Harden tissues excessively & distorts tissue
(1:1)  Used to fix brain tissues for diagnosis differentiation. morphology
of rabies  Preserves Nissl granules and cytoplasmic  Dissolves fat, lipids, myelin
Fixation Time: 1-3 HOURS granules well.  Leads to Polarization (unless very cold temp [-
Carnoy’s Fluid
 Preserves nucleoproteins and nucleic acids. 70C] are used)
 Excellent fixative for glycogen since aqueous  Dissolves acid-soluble cell granules and
solutions are avoided. pigments
 Very suitable for small tissue fragments such
as curettings and biopsy materials.
 Following fixation, tissues may be transferred
directly to absolute alcohol, thereby
shortening processing time.
 Rec for fixing mucopolysaccharides and
Newcomer’s Fixation Time: 12-18 HOURS @ nuclear proteins.
Fluid 3°C  Produces better reaction in Feulgen stain than
Carnoy's fluid.

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 Chapter 3: FIXATION AND FIXATIVES | 10
 Acts both as a nuclear and histochemical
fixative.

Osmium tetroxide (osmic acid)

FIXATIVE DESCRIPTION PURPOSE ADVANTAGES DISADVANTAGES
 Most common chromium-  Recommended for nuclear  It is an excellent fixative for nuclear  It is a poor penetrating agent; hence, is
osmium acetic acid fixative preparation of such sections structures, e.g. chromosomes. applicable only to small pieces of tissues.
used  It permanently fixes fat.  The solution deteriorates rapidly and must be
 Relatively less amount of solution is required prepared immediately before use.
Fixation Time: 24-48 HOURS for fixation (less than 10 times the volume of  Chromic-osmic acid combinations depress the
Flemming’s the tissues to be fixed). staining power of hematoxylin (especially
Solution Ehrlich's hematoxylin).
 It has a tendency to form artifact pigments;
these may be removed by washing the fixed
tissue in running tap water for 24 hours
before dehydration.
 It is very expensive.
 Made up of chromic and  Recommended for cytoplasmic  Same as Flemming’s  Same as Flemming’s
osmic acid structures particularly the
Flemming’s
 Removal of acetic acid from mitochondria
Soluton w/o
the formula serves to
acetic acid
improve the cytoplasmic
detail of the cell

Trichloroacetic acid
FIXATIVE DESCRIPTION PURPOSE ADVANTAGES DISADVANTAGES
 reagent that is used for the  Precipitates proteins.  Poor penetrating agent; suitable only for
precipitation of proteins and nucleic  Its marked swelling effect on tissues serves to small pieces of tissues or bones
acids counteract shrinkage produced by other
Tricholoracetic
 Also used as a decalcifier and fixative fixatives.
Acid
in microscopy.  May be used as a weak decalcifying agent
 Sometimes incorporated into  Its softening effect on dense fibrous tissues
compound fixatives facilitates preparation of such sections.

Acetone
FIXATIVE DESCRIPTION PURPOSE ADVANTAGES DISADVANTAGES

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Acetone  Used at ice cold temperature
ranging from -5°C to 4°C
 Fixation time may vary from
several minutes (for cell
smears, cryostat sections) to
several hours (1-24 hours for
small tissue blocks).
Heat fixation
FIXATIVE DESCRIPTION
 Involves thermal coagulation
of tissue proteins for rapid
diagnosis
 Its use is reserved for the fixation of
cryostat sections or for tissues in
which enzymes have to be preserved.
PURPOSE
 Usually employed for frozen tissue
sections and preparation of
bacteriologic smears
Chapter 3: FIXATION AND FIXATIVES | 11
 It is recommended for the study of water  Produces inevitable shrinkage and distortion
diffusible enzymes especially phosphatases  Dissolves fat
and lipases.  Preserves glycogen poorly
 It is used in fixing brain tissues for diagnosis of  Evaporates rapidly
rabies.
 It is used as a solvent for certain metallic salts
to be used in freeze substitution techniques
for tissue blocks.
ADVANTAGES DISADVANTAGES
 Fixation is better  Produces considerable tissue shrinkage and
 Preserves nuclear and cytoplasmic detail distortion
 Suitable for frozen tissues preparation  Destroys RBC
 Dissolves starch and glycogen
@ohkeye & 楊桂霞