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1186
Medical Laboratory Technology: Volume 3
4. Wash in several changes of distilled water.
5. Wash well in tap water.
6. Counterstain as required, either in van Gieson or neutral red for 5 min.
7. Dehydrate by taking the slide through increasing concentration of alcohol.
8. Clear and mount in DPX.
• Black in colour—calcium deposits
• Background colour—according to counterstain used
• Cell nuclei are red and cytoplasm is pink with neutral red.
StainS fOr micrOOrganiSmS
Tissue sections are sometimes examined for the presence of certain microorganisms with
special reference to bacteria and fungi (systemic). Many of these
however, special stains are employed to make them easier
to identify. Some of these microorganisms may not pick up the routine stain, or they may be
very few in numbers and are almost impossible to nd without special stains. Only a few of
the routinely used methods followed in Indian laboratories will be discussed here: Gram stain
for bacteria, acid fast stain for mycobacterium (tuberculosis and leprosy), silver methenamine
stain for fungi, and Giemsa stain for bacteria and blood parasites. A few commonly used
histological stains to demonstrate the presence organisms in tissues are given in Table 37.3.
Table 37.3 Commonly used histological stains to demonstrate organisms in tissues
Stain
Haematoxylin-eosin (H&E) Viral, Toxoplasma, and fungi
(Aspergillus, Zygomycetes)
Demonstrate cellular response.
Gomori’s methenamine silver nitrate
(GMS)
Periodic acid-Schi (PAS) Fungi, sporotrichosis, Blastmyces,
Acid fast Mycobacteria and Nocardia Mycobacteria
Gram Bacteria, Actinomyces, and fungi Gram reacting bacteria, Actinomyces,
Giemsa Rickesia, Pneumocystis (will stain
Fungi (yeast and mycelial form) Histoplasma, Candida, Aspergillus
and Zygomycetes
internal structure), and fungi
CMV, Herpes, Adenovirus,
Toxoplasma
Blastomyces
Zygomycosis
Coccidiodes
Nocardia
Histoplasma, Pneumocystis, Candida
General Purpose Stain
Giemsa stain
Giemsa stain is widely used as a bacterial stain, for the study of haematological elements, for
bone marrow preparations, for blood parasites. Its use in histological specimens is a modied
version of the original method that gives good results. Most acidophilic cells and eosinophils
stain a similar colour. To achieve a good colour balance, the sections are rst over stained
with Giemsa and then dierentiate in weak acetic acid until there is a general pink cast to the
Laboratory Techniques in Histology
cells. This osets the loss of eosinophilia and gain in basophilia which results upon alcohol
dehydration. Decalcication of tissue with strong acids should be avoided as it results in a
poor colour balance. The only real disadvantage is the length of time required in staining.
• Giemsa stain:
This staining solution is readily available commercially but, to prepare a batch, the
following formula may be use.
■ Mix 7.36 g of Giemsa dry stain in 500 mL glycerol heated to 50°C in a water bath.
■ Leave for 30 min at 50°C with periodic mixing.
■ Allow to cool and add 500 mL methanol (the acetone content is immaterial).
■ Mix and lter.
• Acetic acid (dierentiator): This nal solution is of 1 : 10,000 v/v strength of weak acetic
acid.
A convenient way of preparing the weak solution required in the staining procedure is
to take 1 mL of 1% acetic acid solution (v/v, aqueous; 1 mL acetic acid mixed with 99 mL
of water) and dilute with 99 mL of distilled water. The total dilution = 100 × 100 times =
10,000 times.
1. Section to distilled water.
2. Filter enough Giemsa stain into a Coplin jar lled with distilled water to render the
solution a dark blue colour (0.5–1.0 mL).
3. Stain with the solution for at least 20 min at 56°C until the section is overall dark blue.
4. Rinse in distilled water and dierentiate in weak acetic acid solution (1:10,000 v/v) until
the section is predominantly pink in colour.
5. Rinse in distilled water.
6. Dehydrate the sections by taking through increasing concentration of alcohol.
7. Clear and mount as desired.
1187
• Nuclei—purple
• Azurophilic granules—blue
• Red blood cells—yellow to pink
• Eosinophil granules, red blood cells and other acidophil structures—pink
Bacteria
Gram staining for bacteria
Details of the Gram staining procedure are presented in the chapter on Bacteriology. The
means by which some organisms retain the blue primary dye (Gram-positive) and others
(Gram-negative) lose it upon dierentiation is due to the action of iodine. The exact way in
which iodine inuences dye fastness to dierentiation is not clear. The traditional view is
that iodine combines with the dye and Gram-positive organisms act as a type of mordant or
trapping agent to form a solvent-resistant dye complex. Fixation is not important but initial
freshness of tissue and the use of positive controls are, so that should the test section be
negative, it is an indicator that the solutions and the skill of the operator are beyond reproach.
Both Gram-positive and Gram-negative bacteria are demonstrated in the following technique.
Although over dierentiation with acetone is possible, it is the method of choice for
1188
consistently good results. The technique for smears is similar in principle to that for sections
and the points to remember are that they should either be heat-dried on a hot plate before
staining, or wet-xed smears through xylene and alcohols. Tissue which has been decalcied
in strong acids (or too long in weak acids) will give inconsistent staining.
• Crystal violet stain:
A. Crystal violet solution (0.5%, w/v, in 25% alcohol)
B. Sodium bicarbonate (5%, w/v, in water)
Freshly mix 1 mL of solution A and 5 drops (0.25 mL) of solution B. Leave for 1 min.
• Gram’s iodine solution:
Add 1 g iodine in 2 g potassium iodide plus a few mL of distilled water in a mortar.
Grind until dissolved and make up to a 300 mL volume. KI facilitates dissolving the
iodine in aqueous medium.
• Neutral red (1%, w/v, aqueous)
• Acetone
1. Take the sections to water (take through decreasing strength of alcohol).
2. Stain with crystal violet solution for 2 min.
3. Wash in water.
4. Treat with Gram’s iodine solution for 2 min.
5. Wash, dehydrate quickly, clear and mount in DPX-type mount.
• Blue: Gram-positive bacteria, some fungi
• Red: Gram-negative bacteria and nuclei
• Yellow: Other tissues
Medical Laboratory Technology: Volume 3
Acid fast staining
Tubercle bacilli (Mycobacterium) have a lipid-rich cell wall capable of taking up strong
phenol-dye solutions so that they retain the dye upon subsequent dierentiation in acid or
alcohol. In other words, they are acid-fast and alcohol-fast. Most other organisms lose the dye
and take up the counterstain. It has been shown that acid-fastness of mycobacterial stained
with carbol fuchsin is due to the presence of carboxyl and hydroxyl groups on unsaturated
lipids present in bacterial cells.
The Ziehl-Neelsen procedure is the traditional method of acid-fast staining and is reliable.
The histological appearance in tuberculosis is usually (but not always) typical; even so, the
demonstration of tubercle bacilli is a useful conrmatory procedure.
If potentially tuberculous unxed material for cryostat sectioning is received, full aseptic
procedure should be adopted, that is, use of gloves, gowns, etc. Following sectioning the
slides should be xed in 10% formalin for at least 10 min before staining, and all equipment
including the cryostat interior should be thoroughly sterilized in either formalin or a
glutaraldehyde solution.
• Carbol fuchsin solution:
Dissolve 1 g basic fuchsin (use the coarse granule, not the more puried type specied
for Schi s reagent) in 10 mL ethanol. Dissolve 5 g of phenol in 100 mL of distilled water.
Mix the two solutions together. Store at room temperature and lter before use.
• 1% Acid–alcohol:
Conc. hydrochloric acid, (sp.gr. 1.19) 3 mL
Alcohol, 70% 99 mL
• Methylene blue solution (0.2%, w/v, aqueous)
Laboratory Techniques in Histology
1189
1. Take the sections on the slide to water (pass through decreasing concentrations of
alcohol).
2. Filter on the carbol fuchsin and heat the slide three times until the ‘steam rises’ (over a
period of 10 min), or heat in a Coplin jar at 56°C for 30 min.
3. Wash well with water.
4. Dierentiate in 1% acid-alcohol. This may take 1–10 min.
5. Wash in water for 5–10 min.
6. Counterstain with the methylene blue solution for 30 s.
7. Wash in water.
8. Dierentiate and dehydrate in alcohol until the sections are a weak blue.
9. Clear and mount in a DPX-type mount.
• Tubercle bacilli—magenta
• Background—weak blue
• Erythrocytes—with slight reddish tint; these are a good index against over decolouri-
zation (yellowish orange).
Weak acid-fast staining This method is recommended for delicate organisms such as
Mycobacterium leprae. Compared with tubercle bacilli, the leprosy bacilli are much less
acid-fast and alcohol-fast and their lipid envelope is more easily aected by fat solvents,
diminishing the staining reaction. Initial dewaxing is done in a mixture of a vegetable oil and
xylene. It is important not to over-stain with the methylene blue, as it will not be possible to
remove the excess dye in alcohol.
• Carbol fuchsin solution: As described before.
• Oil-xylene mixture: One part vegetable oil (peanut oil or clove oil) and two parts xylene.
• Acid—alcohol:
Concentrated hydrochloric acid (sp. gr. 1.19) 1 mL
Alcohol (70%) 100 mL
• Methylene blue solution (0.2%, w/v, aqueous)
1. Warm the section and deparanize by placing in the oil mixture. Leave for at least
10 min.
2. Blot dry the oil mixture and wash in water. This step may be repeated, should any
xylene-oil remain on the section.
3. Filter on the carbol fuchsin solution for 20 min at room temperature. Do not heat.
4. Wash.
5. Dierentiate in 1% acid-alcohol for 1 min.
6. Wash well in water and counterstain in weak (0.2%) methylene blue for 5–10 s.
7. Wash, wipe edges, blot dry and clear in xylene. Repeat the bloing-xylene treatment
until the section is clear.
8. Air dry.
9. Mount in a DPX-type mountant.
• Magenta—Leprosy bacilli, red blood cells
• Background—pale blue
1190
Fluorescencetechniquefortuberclebacilliandleprosybacilli Fluorescence demonstration
of tubercle bacilli (and also leprosy bacilli) is superior to the acid-fast staining technique.
But it requires special microscope and some expertise in its preparation. In this technique,
background tissue uorescence is masked by the potassium permanganate treatment. To
stain for leprosy bacilli, avoid alcohol dehydration at the conclusion of the technique and
de-wax initially in a vegetable oil–xylene mixture (as discussed earlier).
• Auramine-rhodamine solution:
Add 1.5 g of auramine O and 0.75 g rhodamine B to 50 mL of distilled water and 75 mL
of glycerol. Mix and add 10 mL phenol liqueed by melting at 56°C. The solution will
keep for up to 2 months. Filter before use.
• Acid–alcohol solution:
■ 0.5% hydrochloric acid in 70% alcohol for tubercle bacilli
■ 0.5% aqueous hydrochloric acid for leprosy bacilli
• Potassium permanganate solution (0.5%, w/v, aqueous)
1. Take the sections to water (pass through decreasing strength of alcohol).
2. Treat the section with preheated (60°C) auramine-rhodamine mixture for 10 min.
3. Wash in water for 2 min.
4. Dierentiate in hydrochloric acid solutions (with or without alcohol) for 2–3 min as
appropriate in dierentiation.
5. Wash in water for 2 min.
6. Treat with the potassium permanganate solution for 1 min.
7. Wash in water for 2 min; blot dry.
8. Dehydrate (omit this step for leprosy bacilli), clear and mount in a DPX-type mountant.
Medical Laboratory Technology: Volume 3
Demonstration of Helicobacter
Helicobacter pylori (formerly assigned to the Campylobacter group of organisms) is an
organism that, since its recognition in 1984, has been rmly linked to gastritis and duodenal
ulceration. It is a Gram-negative bacillus having the distinctive morphology of a stumpy
cured rod. Staining by the conventional Gram staining gives indierent results in histological
materials. The cresyl fast violet method, as described here, is simple and eective.
In gastric biopsies the organisms (Helicobacter pylori) are normally found on the epithelial
surface or in the mucosal glandular folds. It is recommended that a positive control slide is
taken through, if possible, so that the relative colour balance of organisms to background can
be assessed.
Cresyl fast violet technique This is a popular technique and simple to do although
dierentiation of the due requires experience before good results are obtained.
• Cresyl fast (echt) violet (1%, w/v, aqueous)
• Alcohol: 95% and absolute.
1. Section to water.
2. Stain with the cresyl fast violet solution for 15 min.
3. Wash in water and dierentiate in 95% alcohol until the organism can be recognized by
its purple colour. Run a positive control for comparison.
Laboratory Techniques in Histology
4. Rinse in absolute alcohol.
5. Clear and mount as desired.
• Helicobacter—purple
• Background—pale blue-purple
1191
Fungi
Silver methenamine stain
Fungi have lile anity for Gram stain; only a few are acid-fast (e.g., Nocardia), and some of
the fungi do not take the H&E routine stain. Gomori’s methenamine–silver nitrate technique
is routinely used in many laboratories as a general purpose fungal stain. The primary
disadvantage is that the silver precipitation method causes fungi to stain black. In tissue
containing black pigment from other sources such as the lung with carbon pigment, the
organisms may go unidentied. Thus, in the case of lung and pulmonary lymph nodes, a
modied PAS stain is used. When in doubt, both the methods can be adopted regardless of
the type of tissue.
The Gomori’s methenamine silver nitrate stain has been employed for the demonstration
of glycogen and mucin. It is, however, widely used for the demonstration of certain fungi
in tissue, more particularly the systemic fungi and opportunist fungi (Aspergillus fumigatus,
Candida albicans). The method is not specic for fungi and is time-consuming, but rarely fails
to demonstrate any fungi present in tissue. A control should always be taken through to
establish the ecacy of the reagents employed.
The connective tissue is blackened by over impregnation making fungal identication more
dicult. Terminate treatment with the hexamine-silver when the fungi appear a dark brown
colour and the background is still clear, that is, aim at slightly under-impregnating. The
inclusion of the borate gives a nal pH of approximately 8.0.
• Sodium metabisulphite solution (1%, w/v, aqueous)
• Chromium trioxide solution (chromic acid): 5%, w/v, aqueous.
• Silver nitrate-methenamine solution
Stock solution: Hexamine-silver
Mix 100 mL of 3% aqueous hexamine and 5 mL of 5% aqueous silver nitrate. A white
precipitate will form that dissolves on shaking. This solution can be kept for 1–2 months
in the refrigerator.
Working solution:
Step 1: Prepare 5% aqueous sodium tetraborate (fresh).
Step 2: Dilute 2 mL of solution 1 (5% sodium tetraborate) with 25 mL distilled water.
Step 3: Mix and add 25 mL of
• Gold chloride solution (0.1%, w/v, aqueous): 100 mg mixed with 100 mL distilled water. This
solution may be used repeatedly.
• Sodium thiosulphate or hypo solution (5%, w/v, aqueous): 5 g sodium thiosulphate in 100 mL
of water.
• Light green counterstain (0.2% in 0.2% acetic acid)
First prepare 0.2% acetic acid (v/v) by mixing 0.2 mL of glacial acetic acid with 100 mL
of distilled water. To this add 0.2 g (200 mg) of light green, S.F. (yellow)
Light green, S.F. (yellow) 0.2 g
Distilled water 100 mL
Glacial acetic acid 0.2 mL
1192
1. Take both test and positive control sections to water (dip in decreasing strengths of
alcohol).
2. Then treat the sections with chromic acid solution for 1 h.
3. Wash in water.
4. Bleach in the metabisulphite solution for 1 min.
5. Wash in tap water for 5–10 min, then in several changes of distilled water.
6. The working hexamine solution should have been preheated to 56°C using a Coplin jar
in a water bath. Treat the section in this solution at 56°C and examine aer 10–20 min
and aer that at 3 min intervals until the fungi are blackened but the background is
clear. Use plastic forceps to hold the slides.
7. Wash in three changes of distilled water and tone in 0.1% aqueous gold chloride for 3 min.
8. Wash in water, x in sodium thiosulphate for 5 min. Wash again.
9. Counterstain in light green for 1–2 min. Wash in water.
10. Dehydrate, clear and mount as desired.
• Black: Fungi, cellulose, mucins, glycogen, starch
• Pale green: Background
• Old rose: Inner parts of micelle and hyphae
Medical Laboratory Technology: Volume 3
Tissue Parasite Demonstration
In developing countries with warmer climates, demonstration of the presence of tissue
parasites in the sections helps in the diagnosis of parasitic infections. Although H&E
preparation and Giemsa staining may be adequate but special technique may be applied to
show the variegated structure of helminths and protozoa. The one that is particularly useful
is the identication of Leishman-Donovan bodies for the diagnosis of Leishmania donovani
infection (kalazar). These parasites are minute and not easily seen in H&E and conventional
Romanowsky-type staining of tissue sections. Other methods have been discussed in Chapter
22 of Vol. II.
• Leishman stain: This is commercially available from most laboratory suppliers. It is
diluted with buer (pH 4.7) before use.
Laboratory preparation:
■ Rinse out a clean staining bottle with methanol.
■ Add a few dry glass beads.
■ Add 0.75 g of Leishman powder and 500 mL of methanol (CH3OH).
■ Mix well.
■ Label with date. The stain is ready for use the following day. It is important to prevent
moisture from entering the stain during its preparation and storage.
• Weak solution of acetic acid (1:10,000, v/v): As described before (Giemsa).
1. Take sections to distilled water (Rehydrate: pass through decreasing grades of alcohol).
2. Treat with diluted Leishman stain for 10 min or 1–2 h. Note Adjust the staining time
when a new batch of stain is used or the stain has been stored for a long time.
3. Rinse in distilled water and dierentiate in weak (1 in 10,000) acetic acid until the desired
colour balance is achieved.
Laboratory Techniques in Histology
4. Rinse in distilled water thoroughly and then blot dry.
5. Dehydrate quickly, clear and mount as desired.
1193
Staining KitS frOm cOmmerciaL cOmpanieS
Acid-Fast Bacillus (AFB) light green stain
Staining kits are now available in the market. Contact your local vendors. The stain is intended
to identify acid-fast bacteria, such as Mycobacterium, in tissue sections on the ArtisanTM Link
and ArtisanTM Link Pro Staining Systems. Application of Carbol Fuchsin stains acid-fast
bacteria red, followed by decolourization of all tissue elements except the acid-fast bacteria.
A Light Green counterstain is then applied to impart a green colour to all background tissue
elements.
Colloidal iron stain
Prepared colloidal iron stain kit is available in local market. It is used to identify carboxylated
and sulfated mucopolysaccharides and glycoprotein mucin in tissue sections. Colloidal ferric
ions are, at a low pH, absorbed principally by carboxylated and sulfated muco-substances
staining a dark blue. A counterstain of Nuclear Fast Red stains the nuclei and cytoplasm light
pink.
Giemsa stain
Prepared Giemsa stain is now available in the market. It is used to permit dierentiation of
cells present in sections cut at 2–3 mm. Polychromatic stains are used as
routine nuclear and cytoplasmic stains for bone marrow biopsy sections because of the colour
range of the cytoplasmic staining and the dierentiation of hematopoietic cells achieved.
• Nuclei: blue
• Eosinophils: bright pink
• Leucocytes: shades of pink, gray, or blue depending on cell type and development.
This stain is not recommended for bone marrow smears.
Gram yellow stain
Gram yellow staining kit is available in the market. The stain is intended to identify
in tissue sections. Gram Yellow Stain
Kit is used to identify two distinct groups of microorganisms in tissue sections. Those that
retain primary dye (Crystal Violet) are called Gram-positive. Those that lose the primary dye
during decolourization are called Gram-negative. Mechanisms of Gram-positive organisms
retaining the primary stain and Gram-negative organisms losing the primary stain are the
result of the chemistry and structure of the organism’s cell walls.
• Gram-positive organisms: dark blue
• Gram-negative organisms: light pink to magenta
• Background: yellow
Grocott’s Methenamine Silver (GMS) eosin stain (available as kit)
The stain is intended to identify and Pneumocystisjirovecii (formerly
known as carinii) in tissue sections on the ArtisanTM Link and ArtisanTM Link Pro Staining
Systems.
Fungi and Pneumocystis jirovecii are stained black while other tissue elements are stained
pink. This stain is not recommended for cytology specimens.
1194
Medical Laboratory Technology: Volume 3
Jones’ Basement Membrane (PAS-M) H&E stain kit
The stain is used to identify basement membranes, specically glomerular and tubular
basement membranes in renal tissue sections cut at 2 mm on the ArtisanTM Link and
ArtisanTM Link Pro Staining Systems.
• The Bowman’s capsule: black
• Inner basement membrane: black to gray
• Nuclei: blue, collagen, cytoplasm
• Other tissue elements: light green
Jones’ Basement Membrane (PAS-M) light green stain kit
This stain is used to identify basement membranes, specically glomerular and tubular
basement membranes in renal tissue sections cut at 2 mm.
• The Bowman’s capsule: black
• Inner basement membrane: black to gray
• Other tissue elements: pink
Orcein stain kit
Orcein staining technique is used to identify viral inclusion bodies of hepatitis B surface
antigen (HBsAG) and copper-associated proteins in tissue sections. Virus particles inside
host cells are called A hepatitis B virus lies on the surface of the virus
particle. can be detected by using the Orcein staining method. The antigen appears
as ne granules either diusely spread throughout the cytoplasm or concentrated in the
cytoplasm peripheral to the sinusoid space. Copper-associated proteins when in excessive
pathologic amounts, such as Wilson’s disease and some forms of cirrhosis can be detected by
using the Orcein staining method.
• HBsAG/elastin bers: dark reddish brown
• Copper-associated protein: dark red-purple
• Background: pale pink
Special Stains
Acid-Fast Bacteria (AFB) stain kit
AFB stain kit is intended to identify acid-fast bacteria, such as Mycobacterium, in tissue
sections on the ArtisanTM Link and ArtisanTM Link Pro Staining Systems. Application of
Carbol Fuchsin stains acid-fast bacteria red, followed by decolourization of all tissue elements
except the acid fast bacteria. A Methylene Blue counterstain is then applied to impart a blue
color to all background tissue elements.
Alcian Blue/PAS stain kit
This stain is intended to identify acidic and neutral mucins in tissue sections. It clearly
separates the acidic and neutral mucins by colour and can be used to distinguish all mucins
in tissue sections. Alcian Blue pH 2.5 stains the acid mucin blue while the Schi's reagent
stains the neutral mucins pink to red. Mixtures of the two mucins will appear purple due to
the positive reactions with both Alcian Blue and Schi's reagent. This stain can be used with
the digestive enzyme, ArtisanTM Alpha-Amylase (Code AR171), for the demonstration of
glycogen in tissue.
Alcian Blue/PAS/Hematoxylin stain kit
Alcian Blue/PAS/Hematoxylin stain kit is intended to identify acidic and neutral mucins in
tissue sections. It clearly separates the acidic and neutral mucins by color and can be used
Laboratory Techniques in Histology
to distinguish all mucins in tissue sections. Alcian Blue pH 2.5 stains the acid mucin blue
while the Schi's reagent stains the neutral mucins pink to red. Mixtures of the two mucins
will appear purple due to the positive reactions with both Alcian Blue and Schi’s reagent. A
hematoxylin counterstain is then applied to impart a blue/black colour to the nuclei.
1195
Congo red stain kit
This stain is used to identify sections. Amyloid is an abnormal protein
product that can be found in various pathologic conditions. This stain demonstrates amyloid
in pink to dark pink with light (bright eld) microscopy or the characteristic apple green
birefringence with polarized light. Mayer’s Hematoxylin is used as a counterstain.
Elastic stain kit
Elastic stain kit is used to identify It utilizes Alcoholic
Hematoxylin, Ferric Chloride and Lugol’s Iodine solutions to stain elastin bers, van Gieson’s
solution is added to dierentiate collagen from elastin. Elastin bers are stained black while
remaining tissue elements are stained as follows:
• Nuclei: blue/black
• Collagen: red
• Other tissue elements: yellow
Feulgen stain kit
This stain is intended to identify deoxyribonucleic acid (DNA) in tissue sections. The DNA
is stained magenta with Schi's reagent, and the stained DNA is contrasted against a green
background.
Giemsa stain kit
As already discussed before.
Gomori’s blue trichrome stain kit
Gomori’s blue trichrome stain kit is used to identify collagen bers in liver and kidney tissue
sections. Gomori’s Blue Trichrome Stain is oen used to demonstrate increased collagen
deposition that is associated with replacement of functional tissue by scar tissue. This stain is
useful in the assessment of cirrhosis of the liver in which thickened collagen replaces normal
tissue causing liver dysfunction. In Gomori’s one-step procedure, the collagen and nuclei are
stained blue; cytoplasm, erythrocytes, and brin are stained pink to red.
Gomori’s green trichrome stain kit
Gomori’s green trichrome stain kit is used to identify collagen bers in liver and kidney tissue
sections on the ArtisanTM Link and ArtisanTM Link Pro Staining Systems. Gomori’s Blue
Trichrome Stain is oen used to demonstrate increased collagen deposition that is associated
with replacement of functional tissue by scar tissue. This stain is useful in the assessment
of cirrhosis of the liver in which thickened collagen replaces normal tissue causing liver
dysfunction. In Gomori’s one-step procedure, the collagen and nuclei are stained green;
cytoplasm, erythrocytes, and brin are stained pink to red.
Gram stain kit
Gram stain kit is used to identify two distinct groups of microorganisms in tissue sections.
Those that retain primary dye (Crystal Violet) are called Gram-positive. Those that lose the
primary dye during decolorization are called Gram-negative. Mechanisms of Gram-positive