FIXATIVE COMPOSITION ADVANTAGE DISADVANTAGE

Aldehyde 40% formalin diluted 1. Fast 1. Irritating fixatives

- Routine - 1:10 (10%) penetration 2. Soft fixative
paraffin section - 1:20 (5%) 2. Cheap and 3. Tissue shrinkage
- Electron stable 4. Prolonged
microscopy 3. Compatible fixation may lead
- Enzyme studies with many to bleaching
stains
Formaldehyde 4. Does not over
harden tissue
5. Preserves fat,
mucin,
glycogen,
proteins.
6. Nervous tissue
preservation
10% Formol-Saline 40% formalin + 1. Even tissue 1. Same as
10%NaCl penetration formaldehyde
2. Minimum 2. Slow fixative
shrinkage and 3. Reduce
distortion metachromatic
3. Ideal for most reaction
staining 4. Acid stain less
technique
4. Preserves fat,
mucin,
glycogen,
proteins.
5.
10% NBF 40% Formalin + 1. Same as 1. Time consuming
NaH2PO4 +Na2HPO4 formol-saline 2. Reduce PAS
2. Prevents reaction of mucin
precipitation 3. Inert towards
of acid lipids
formalin
pigments
3. Best for iron
pigment
 containing
tissue

Formol corrosive/ 40% Formalin + sat. Aq 1. Rapid 1. Mercuric chloride

Formol Sublimate HgCl2 penetration of deposits
small pieces of 2. Not for frozen
tissue section
2. Minimum 3. Inhibits
shrinkage and determination of
hardening extent tissue
3. Excellent for decalcification
many staining
procedures
4. Does not
require
washing
5. Lipid fixative
Gluteraldehyde - 2.5% / 2-4 1. More stable 1. More expensive
- Routine light hours for small effect 2. Reduced PAS
microscopy tissue/ needle 2. Preserve reaction of mucin
- Electron biopsy plasma
microscopy - 4.0%/ 6-8-24 protein and
- Enzyme hrs – for larger cellular
histochemistry tissue (<4mm) structure
better
3. Less irritating
Metallic Fixative: Chromate

FIXATIVE COMPOSITION ADVANTAGE DISADVANTAGE

1. Chromic Acid 1-2 % K2Cr207 1. Precipitates 1. Easily

all protein decomposed
and upon prolonged
adequately standing
2. Strong 2. Add strong
oxidizing reducing agent to
agent chrome fixatives
2 3% K2Cr207 3% K2Cr207 1. Preserves 1. pH requirement
lipid
2. Preserves
mitochondria
at pH 4.5 –
5.2
3. Regaud’s 3% K2Cr207 + 40% 1. Penetrates 1. Tissue must be
- For formaldehyde tissue well <2-3 mm
demonstration 2. Hardens 2. Deteriorates and
of chromatin more rapidly darkens
mitochondria, than Orth’s
mitotic figures
, golgi bodies,
RBC and
colloid
containing
tissue
4. Orth’s Fluid 2.5% K2Cr207 + 1. Demonstrate Same as Regaud Fluid
Na2so4 + 40% Rickettsiae
Formaldehyde and other
bacteria
2. Precerve
Myelin
Metallic Fixative: Lead
Lead Fixatives – are generally for Acid Mucopolysaccharide
ex. Umbilical cord/ Wharton’s Jelly

FIXATIVE COMPOSITION ADVANTAGE DISADVANTAGE

1. Lead Acetate 4% Lead Acetate 1. Fixes 1. Prolonged
- For Acid connective standing may
Mucopolysaccharide Tissue lead to
formation of
lead
carbonate

Metallic Fixative: Mercuric Chloride

Mercuric Chloride – may produce black granular deposits on tissues

FIXATIVE COMPOSITION ADVANTAGE DISADVANTAGE

Mercuric Chloride HgCl2+ K2Cr207 + 1. More 1. Prolong
Na2SO4 + H20 convenient standing may
and easier to lead to
use
deterioration
Zenker’s Fluid Stock solution + 1. For trichrome 1. Poor penetration
Glacial HAC staining 2. Deteriorates
2. Acts as upon addition
mordant HAC
Heidenhien’s HgCl2 Trichloro- 1. Minimum 1. RBC
SuSa HAC+Glacial shrinkage preservation
HAC + 40% and
2. Mercuric
Formaldehyde hardening
2. For silver Chloride
impregnation deposits
 3. Use of
Welgert’s stain
is not possible

PICRIC ACID FIXATIVE

- Highly explosive when dry
- Will produce excessive staining of tissue
- Picrate’s are form upon protein ; precipitates are soluble in water; hence
tissues must be first rendered insoluble by direct immersion in 70% ETOH
- Picrate fixative must never be washed in water before dehydration

FIXATIVE COMPOSITION ADVANTAGE DISADVANTAGE

Picric acid 1% Picric acid 1. Stable and 1. RBC lysis

- Excellent for does not 2. Not for frozen
demonstration deteriorate section
of glycogen rapidly 3. Alters and
2. Used for dissolve lipid
trichrome
methodand
aniline stain
Bouin’s Solution Sat. Sol. Of Picric Acid 1. Minimal 1. Limited to small
+ 40% formaldehyde + distortion on fragments
glacial HAC microanatomical 2. RBC lysis
structure 3. Not suitable for
2. Yellow stains fixing kidneys
useful when
handling
3. Brilliant staining
of tissue
4. Preserves
glycogen
Brasil’s Alc, Picric acid + 37% 1. Less messy and 1. Tedious process
Picroformol Fixative Formaldehyde more convenient
- Excellent for
demonstration
of glycogen