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1. TITLE: Special Stains Overview.
2. DEFINITION :
Special staining is performed to visualize selected tissue elements, entities and microorganisms. Based on classical dye staining
methods, special stains technique provide valuable information in the evaluation of numerous abnormal or disease conditions. The
following special stains are of high quality, and they satisfactorily demonstrate (on each day of use), the tissue components or
organisms for which they were designed.
a) Acid fast organisms
b) Iron
c) Bacteria
d) Elastic tissue
e) Fungi
f) Mucin
g) Connective tissue
h) Myelin
i) Nerve fibers
j) Glycogen
k) Reticulin fibers
l) Amyloids
3. POLICY :
Nil.
4. PROCEDURE :
4.1. SAFETY.
4.1.1. All specimens should be handled with care as if they are infectious.
4.1.2. All specimens should be handled with care as if they are infectious.
4.1.3. Always wear Personal Protection Equipment’s (PPE).
4.1.4. For more safety information, refer to MSDS.
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4.2. QUALITY CONTROL.
4.2.1. Always run a known positive control with the test.
4.2.2. Positive tissue controls assess the performance of the stain. Special stains are performed on sections of control tissue known
to contain components specific to each special stain.
4.2.3. Verification of tissue used as a positive control must be performed and documented before being used with clinical
specimens.
4.2.4. Document the evaluation of positive control tissue block on Lab.Med.FRM-HIST-021.
4.2.5. Document daily QC on Lab.Med.FRM-HIST-032.
4.2.6. Check expiry date of the kit prior to run.
4.2.7. Perform maintenance of the machines used for special stains.
4.3. INSTRUMENT:
4.3.1. Dako Artisanlink Autostainer
(Refer to IPP 14.HIST 014 Dako Autostainer for Special stains).
5.
NO SPECIAL INTENDED USE PRINCIPLE CLINICAL CONTROL RESULTS
STAIN SIGNIFICANCE
1. Bacteria of the genus Acid Fast Bacillus A tissue containing Acid fast bacilli: Red
Acid Fast Demonstration of mycobacterium have stain is intended for acid- fast organism. Nocardia Filaments:
acid-fast the unique property use as a qualitative Red
Mycobacteria such as of being “acid fast” histologic stain to Leprae bacilli: Red
M.Tuberculosis, M. with selectively Back ground: Blue
Leprose,M. Kansasii, certain stain. The term demonstrate
M Ayium. acid fact refer to the mycobacterium
ability if the bacterium Tuberculosis and
to retain a dye that other acid-fast
washes organisms or
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out of all other tissue components in
element when the tissue section.
section is exposed to
a dilute acid solution.
Alcian blue is a water
It stains basophiles, Umbilical Acid
2. Alcian Blue – soluble amphoiteric
cartilae,, Mucopolysaccharides:
copper phthalocyanin
Evaluation of Acid mucins and Cord
mucopolysaccharides Turquoise Blue
dye that stain by salt
mucopolysaccharides
and
linkage with the acidic
glycosaminoglycans
groups of acid
mucopolysaccharides.
Alcian Blue /
Differentiation of This is a combination Non-small cell lung Umbilical Cord or · Acid
3. PAS
neutral and acidic of two standard cancer (NSCLC) can Appendix mucopolysaccharides:
mucopolysaccharides techniques: Periodic be classified into Blue
Acid-Schiff and several histological
· Neutral
Alcian Blue subtypes, most
polysaccharides:
commonly
pH 2.5 . Magenta
adenocarcinoma
(LADC) or
· Cartilage, ground
squamous cell
substance, epithelial
carcinoma (SqCC).
mucins: Shades of
With the
purple to very deep
introduction of
blue
targeted therapies
that can result in
· Cell bodies of fungi:
dramatically different
Red to purple
outcomes based on
subtype, the
· Mucoid capsule of
importance of
Cryptococcus: Blue
accurate
classification has
been amplified.
Limited availability
of tissue from needle
core biopsies has
challenged routine
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challenged routine
diagnosis,
particularly for
poorly differentiated
samples. Studies
have shown that
histochemical stains
Alcian blue-PAS are
useful in lung cancer
diagnosis
Congo red stain is a Congo red stain is A tissue section With normal light
4. Amyloid
modification of the used as a qualitative containing amyloid microscopy:
(Congo Red)
Demonstration of
Highman's technique. histologic stain to
Amyloid in tissue Cut on 5 microns Amyloid Pink to red.
The staining reaction selectively
sections
is based on the demonstrate
Nuclei Blue
application of congo AMYLOID in tissue
red which stains the sections
· With polarized light
pattern of a typical
microscopy: Under
proteins (Amyloid).
polarized light to pink
The B-pleated sheets
areas change to green
of amyloid are
suitable in size and
as the filter is rotated.
shape to
The background is
accommodate the
dark.
congo red molecules,
which are held in the
lattice work of B-
pleated sheets.
Birefringence is an
intrinsic property of
the amyloid fibril
congo red complex.
Without both the red
staining congo red
and the apple green
birefringence under
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