Difference between revisions of "Stains"

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This article deals with '''stains'''. H&E isn't the only stain out there...
[[Image:Corpora amylacea high mag.jpg|thumb|300px|[[Hematoxylin and eosin stain]] of benign [[prostate gland]].]]
This article deals with '''stains'''. The H&E stain is only one of many stains that are used in pathology.


==Where to start...==
Non-H&E stains are often referred to as '''''special stains'''''.
===Principles===
 
=Where to start...=
==Principles==
When considering additional (i.e. special) stains one should (in order) do the following:<ref>LAE. 13 July 2010.</ref>
When considering additional (i.e. special) stains one should (in order) do the following:<ref>LAE. 13 July 2010.</ref>
# Make sure one has exhausted the clinical history; history is considered the best special stain.
# Make sure one has exhausted the clinical history; history is considered the best special stain.
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# Molecular testing, electron microscopy.
# Molecular testing, electron microscopy.


===Common stains===
==Common stains==
# [[H&E stain]].
# [[H&E stain]].
# [[PAS stain]].
# [[PAS stain]].
# [[PAS-D stain]].
# [[PAS-D stain]].
# [[AFB stain]].
# [[AFB stains]], e.g. [[Ziehl-Neelsen stain]].
# [[Congo red]].
# [[Congo red]].
# [[GMS stain]].
# [[GMS stain]].
# [[Gram stain]].
# [[Gram stain]].


==Immunohistochemistry==
=Immunohistochemistry=
{{main|Immunohistochemistry}}
{{main|Immunohistochemistry}}
===General===
==General==
*Abbreviated ''IHC''.
*Abbreviated ''IHC''.


===Interpretation===
==Interpretation==
Simple version:
Simple version:
*Positive is (usually): ''brown''.
*Positive is (usually): ''brown''.
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*The edge of the tissue may have light staining - ''edge effect''.
*The edge of the tissue may have light staining - ''edge effect''.
*If everything is brown... suspect that it didn't work.
*If everything is brown... suspect that it didn't work.
*In some situations you're blessed with an ''internal control'', e.g. in renal tumours CD10 will stain [[RCC]] and the ''proximal tubule'', in GISTs - CD117 the mast cells are positive.
*In some situations you're blessed with an ''internal control'', e.g. in renal tumours CD10 will stain [[RCC]] and the ''proximal tubule'', in GISTs - CD117 the [[mast cell]]s are positive.


==Work-up of infection==
=Work-up of infection=
It often not possible to be definitive by staining.<ref>{{cite journal |author=Woods GL, Walker DH |title=Detection of infection or infectious agents by use of cytologic and histologic stains |journal=Clin. Microbiol. Rev. |volume=9 |issue=3 |pages=382-404 |year=1996 |month=July |pmid=8809467 |pmc=172900 |doi= |url=http://cmr.asm.org/cgi/pmidlookup?view=long&pmid=8809467}}</ref>
It often not possible to be definitive by staining.<ref>{{cite journal |author=Woods GL, Walker DH |title=Detection of infection or infectious agents by use of cytologic and histologic stains |journal=Clin. Microbiol. Rev. |volume=9 |issue=3 |pages=382-404 |year=1996 |month=July |pmid=8809467 |pmc=172900 |doi= |url=http://cmr.asm.org/cgi/pmidlookup?view=long&pmid=8809467}}</ref>


Basic panel:
Basic panel:
*Gram stain - for bacteria.
*Gram stain - for bacteria.
*GMS stain - fungal stain.
*GMS stain - [[fungi|fungal]] stain.
*PAS (''or'' PAS-D) - fungal stain.
*PAS (''or'' PAS-D) - fungal stain.


===Fungi===
==Fungi==
{{main|Microorganisms}}
{{main|Fungi}}
 
Fungi are a type of [[microorganisms]].  They are seen by pathologist every once in a while.


==Specific stains==
=Specific stains=
What follows is a big list... of stains.
What follows is a big list... of stains.


==Haematoxylin and eosin stain==
==Haematoxylin and eosin stain==
===General===
*Abbreviated ''H&E''.
*Abbreviated ''H&E''.
*Standard bearer in most pathology departments.
{{Main|Hematoxylin and eosin stain}}
 
===Intepretation===
*Blue (haematoxylin) = nucleus.
*Pink (eosin) = cytoplasm.


==Haematoxylin phyloxin saffron stain==
==Haematoxylin phyloxin saffron stain==
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===Interpretation===
===Interpretation===
*Haematoxylin = blue -- stains nucleus.
*Haematoxylin = blue -- stains nucleus.
*Phyloxin = pink.
*Phyloxin = pink -- stains muscle and cytoplasm.
*Saffron = yellow -- stains collagen.
*Saffron = yellow -- stains collagen.
*An alternative to H&E stain.
*An alternative to H&E stain.
**Fibrosis is easier to see on HPS than H&E... as one can see the collagen.
**Fibrosis is easier to see on HPS than H&E... as one can see the collagen.


====Images====
<gallery>
Image:Perineural_invasion_prostate_high_mag.jpg | Perineural invasion - prostate - HPS stain (WC)
Image:Meningioma_high_mag.jpg | Meningioma - HPS stain (WC)
Image:Endoneurial_fibrosis_-_very_high_mag_-_cropped.jpg | Endoneurial fibrosis - HPS stain (WC)
</gallery>
==Periodic acid Schiff stain==
==Periodic acid Schiff stain==
*Abbreviated ''PAS''.
*Abbreviated ''PAS''.
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===Utility===
===Utility===
*Stains - basement membranes, fungi, glycogen, (neutral) mucin.  
*Stains - lipofuscin,<ref name=pmid5463681 >{{cite journal |author=Kovi J, Leifer C |title=Lipofuscin pigment accumulation in spontaneous mammary carcinoma of A/Jax mouse |journal=J Natl Med Assoc |volume=62 |issue=4 |pages=287–90 |year=1970 |month=July |pmid=5463681 |pmc=2611776 |doi= |url=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2611776/pdf/jnma00512-0077.pdf}}</ref> basement membranes, fungi, glycogen, (neutral) mucin.  


===Interpretation===
===Interpretation===
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*Blue = nuclei.
*Blue = nuclei.


Image: [http://commons.wikimedia.org/wiki/File:Metanephric_adenoma_high_mag.jpg Metanephric adenoma - PAS (WC)].
Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF]</ref>
====Image====
<gallery>
Image:Metanephric_adenoma_high_mag.jpg | Metanephric adenoma - PAS (WC)
File:Liver biopsy of glycogen storage disorder PAS positive.jpg | Liver biopsy with glycogen storage disease - PAS (WC)
File:Esophageal candidiasis (2) PAS stain.jpg | Esophageal candidiasis - PAS (WC)
</gallery>
 
==Periodic acid Schiff fungal stain==
*Abbreviated ''PASF''.
 
===Primary application===
*Look for [[fungi|fungal organisms]].
 
===Interpretation===
*Light purple = fungi.
*Light green = background.
*Washed-out light purple = Gram positive bacilli.
 
Note:
*This is much improved over the ''PAS'' in the context of skin, as the background is similar to the fungal organisms.


Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF]</ref>
<gallery>
Image:Fungal_infection_-_PASF_-_very_high_mag.jpg | Fungal organisms. PASF stain.
</gallery>


==Periodic acid Schiff with diastase==
==Periodic acid Schiff with diastase==
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===General===
===General===
*Diastase digests the glycogen.
*Diastase digests the glycogen.
*"PAS diastase resistant"<ref name=pmid6189389>{{Cite journal  | last1 = Qizilbash | first1 = A. | last2 = Young-Pong | first2 = O. | title = Alpha 1 antitrypsin liver disease differential diagnosis of PAS-positive, diastase-resistant globules in liver cells. | journal = Am J Clin Pathol | volume = 79 | issue = 6 | pages = 697-702 | month = Jun | year = 1983 | doi =  | PMID = 6189389 }}</ref> ''implies'' PASD +ve and [[PAS stain|PAS]] +ve.


===Use===
===Use===
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Notes: <Ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF]</ref>
Notes: <Ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF]</ref>
===Interpretation===
*Light purple = fungi.
*Light blue/pink = background. ???


==Gomori methenamine-silver stain==
==Gomori methenamine-silver stain==
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===Use===
===Use===
*Useful for fungi.  
*Useful for fungi.  
**Pneumocystis jirovecii - cause of ''PCP'' - pneumocystic pneumonia.
**Pneumocystis jirovecii - cause of [[pneumocystis pneumonia]] (PCP).
**Histoplasma - cause of histoplasmosis.
**Histoplasma - cause of [[histoplasmosis]].
***Histoplasma = black, round balls.
***Histoplasma = black, round balls.


Image: [http://commons.wikimedia.org/wiki/File:Histoplasma_in_granuloma_gms.jpg GMS showing histoplasma (WC)].
====Image====
 
<gallery>
Image:Histoplasma_in_granuloma_gms.jpg | GMS showing histoplasma (WC/Nephron)
</gallery>
==Acid-fast bacilli stains==
==Acid-fast bacilli stains==
*Abbreviated: ''AFB''.
*Abbreviated: ''AFB''.


There are several AFB stains:
There are several AFB stains:
*Ziehl-Neelson stain - used to look for Mycobacterium tuberculosis.
*Ziehl-Neelson stain - used to look for ''[[Mycobacterium tuberculosis]]''.
*Fite stain - used to look for Mycobacterium laprae.<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF]</ref>
*Fite stain - used to look for ''[[Mycobacterium leprae]]''<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF]. Accessed on: 19 May 2011.</ref> and preferred stain for ''[[Mycobacterium avium complex]]''.<ref name=pmid21327589>{{cite journal |authors=Ilyas S, Youssef D, Chaudhary H, Al-Abbadi MA |title=Myocbacterium-avium intracellulare associated inflammatory pseudotumor of the anterior nasal cavity |journal=Head Neck Pathol |volume=5 |issue=3 |pages=296–301 |date=September 2011 |pmid=21327589 |pmc=3173547 |doi=10.1007/s12105-011-0248-0 |url=}}</ref>
*Auramine rhodamine.
*Auramine-rhodamine stain.


===Ziehl-Neelsen stain===
===Ziehl-Neelsen stain===
*Most popular acid-fast bacilli stain.
*Most popular acid-fast bacilli stain.
*Stains other mycobacteria -- not specific for tuberculosis.
*Stains other mycobacteria -- not specific for tuberculosis.
**Stains nocardia.<ref>[http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html]</ref>
**Stains ''Nocardia''.<ref>URL: [http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html]. Accessed on: 19 May 2011.</ref>


====Image====
<gallery>
Image:Mycobacterium_tuberculosis_Ziehl-Neelsen_stain_02.jpg | ZN stain. (WC/CDC)
</gallery>
===Fite stain===
===Fite stain===
Interpretation:
Interpretation:
*Red = AFB.
*Red = AFB.
*Blue = background.
*Blue = background.
===Auramine-rhodamine stain===
*Fluorescent stain.
====Image====
<gallery>
Image:Cryptosporidium_parvum_auramine-rhodamine_labeled.jpg | AR stain. (WC/CDC)
</gallery>
===Kinyoun stain===
*Another AFB stain<ref name=pmid7536216>{{Cite journal  | last1 = Kehl | first1 = KS. | last2 = Cicirello | first2 = H. | last3 = Havens | first3 = PL. | title = Comparison of four different methods for detection of Cryptosporidium species. | journal = J Clin Microbiol | volume = 33 | issue = 2 | pages = 416-8 | month = Feb | year = 1995 | doi =  | PMID = 7536216 }}</ref> - useful for [[cryptosporidiosis]] and [[microsporidiosis]].<ref name=pmid9003613>{{Cite journal  | last1 = Ignatius | first1 = R. | last2 = Lehmann | first2 = M. | last3 = Miksits | first3 = K. | last4 = Regnath | first4 = T. | last5 = Arvand | first5 = M. | last6 = Engelmann | first6 = E. | last7 = Futh | first7 = U. | last8 = Hahn | first8 = H. | last9 = Wagner | first9 = J. | title = A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens. | journal = J Clin Microbiol | volume = 35 | issue = 2 | pages = 446-9 | month = Feb | year = 1997 | doi =  | PMID = 9003613 }}
</ref>


==Congo red stain==
==Congo red stain==
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**Mnemonic: ''CRAP'' = congo red amyloid protein.
**Mnemonic: ''CRAP'' = congo red amyloid protein.
*An alternate stain for amyloid is ''Thioflavin T''.
*An alternate stain for amyloid is ''Thioflavin T''.
Note:
*[[Cutting|Thick sections]] (~10 micrometers) are considered a requirement for the stain to work properly.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/congo_red_bennhold.htm http://www.ihcworld.com/_protocols/special_stains/congo_red_bennhold.htm]. Accessed on: 26 January 2012.</ref>
**If the section is too thin... it doesn't work.


===Interpretation===
===Interpretation===
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Ref.:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF]. Accessed on: 4 December 2010.</ref>
Ref.:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF]. Accessed on: 4 December 2010.</ref>


Image: [http://commons.wikimedia.org/wiki/File:Cerebral_amyloid_angiopathy_-_very_high_mag.jpg Congo red stain - cerebral amyloid angiopathy (WC)].
====Image====
<gallery>
Image:Cerebral_amyloid_angiopathy_-_very_high_mag.jpg | Congo red staining in [[cerebral amyloid angiopathy]]. (WC)
</gallery>
 
==Thioflavin T stain==
===Use===
*Used to look for [[amyloid]].
 
===Interpretation===
*Amyloid = green.
 
Image: [http://inano.au.dk/research/annual-reports/annual-report-2004/7-cases/protein-fibrils/ Amyloid (inano.au.dk)].


==Gram stain==
==Gram stain==
Use:
===Use===
*"It is useless for finding bacteria."<ref>SMH handout.</ref>
*"It is useless for finding bacteria."<ref>St. Michael's Hospital - Stains Handout.</ref>
**I suppose this is as you find 'em on H&E... or microbiology finds 'em.
**If they are to be seen... they'll be visible on H&E.
 
Note:
*Microbiology is better at finding organisms than pathology.
**They have one significant advantage -- if a small amount of bugs are present... they grows into a large (obviously visible) colony.
 
====DDx for common patterns====
A short list of bacteria and their characteristics:<ref>URL: [http://www.atsu.edu/faculty/chamberlain/Website/pnebact.htm http://www.atsu.edu/faculty/chamberlain/Website/pnebact.htm]. Accessed on: 7 May 2013.</ref>
{| class="wikitable sortable"
! Shape\Gram stain
! Positive
! Negative
! Variable or negative
|-
| Bacilli
| Clostridium difficile, Bacillus anthracis, Nocardia spp.
| Escherichia coli, [[Helicobacter pylori]], Yersinia pestis, Hemophilus influenzae
| [[Tuberculosis|Mycobacterium tuberulosis]], Legionella pneumophila<ref>URL: [http://meded.ucsd.edu/isp/1999/CAP/legion.html http://meded.ucsd.edu/isp/1999/CAP/legion.html]. Accessed on: 7 May 2013.</ref>
|-
| Cocci
| Streptococcus pneumoniae, Staphylococcus aureus
| Neisseria meningitidis, Moraxella catarrhalis
|
|}


===Interpretation===
===Interpretation===
*Purple (or blue) = Gram positive organisms.
*Purple (or blue) = Gram positive organisms.
*Red = Gram negative organisms, nuclei
*Red = Gram negative organisms, nuclei.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF]. Accessed on: 7 May 2013.</ref>
*Yellow = background.
*Yellow = background.


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*Memory device: '''p'''urple = '''p'''ositive.
*Memory device: '''p'''urple = '''p'''ositive.


Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF]</ref>
====Images====
 
<gallery>
Image:Gram_stain_01.jpg | Gram positive cocci. (WC)
Image:Gram_Stain_Anthrax.jpg | Gram positive rods - anthrax. (WC)
</gallery>
==Luxol fast blue stain==
==Luxol fast blue stain==
*Abbreviated ''LFB''.
*Abbreviated ''LFB''.
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**Lack of blue (where it ought to be) = demyelination.
**Lack of blue (where it ought to be) = demyelination.
*Purple = nerve cell (e.g. neuron).
*Purple = nerve cell (e.g. neuron).
*Neutrophils = pink.
*[[Neutrophil]]s = pink.


Image: [http://en.wikipedia.org/wiki/File:Globus_pallidus_and_putamen_-_very_low_mag.jpg Globus pallidus and putamen - H&E-LFB (WC)].
Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF]</ref>


Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF]</ref>
====Image====
<gallery>
Image:Globus_pallidus_and_putamen_-_very_low_mag.jpg | Globus pallidus and putamen - H&E-LFB. (WC)
File:LFB_CNS_cortex_supratentorial.jpg | Normal cortex - LFB only. (WC/jensflorian)
File:LFB_CNS_cortex_grey-white_matter_junction.jpg | White-grey matter junction - LFB. (WC/jensflorian)
</gallery>


==Giemsa stain==
==Giemsa stain==
===Use===
===Use===
*Useful for finding mast cells.
*Useful for finding [[mast cell]]s.
*Useful for finding donovan bodies and leishmania.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: April 6, 2009.</ref>
*Useful for finding ''[[Donovan bodies]]'' and ''[[Leishmania]]''.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: April 6, 2009.</ref>


===Interpretation===
===Interpretation===
*Tissue is light blue/green.
*Tissue is light blue/green.
*Goblet cells are purple.<ref>URL: [http://www.kennedy.ox.ac.uk/facilities/histology/histology-information http://www.kennedy.ox.ac.uk/facilities/histology/histology-information]. Accessed on: 17 August 2015.</ref>
Image:
*[http://path.upmc.edu/cases/case196/images/figure12.jpg Giemsa - colon (amser.org)].<ref>URL: [http://amser.org/index.php?P=AMSER--ResourceFrame&resourceId=6018 http://amser.org/index.php?P=AMSER--ResourceFrame&resourceId=6018]. Accessed on: 17 August 2015.</ref>


==Reticulin stain==
==Reticulin stain==
===Use===
===Use===
*Liver biopsy, medical.
*Liver biopsy, medical.
**Demonstrates the reticular fibers (in cirrhosis the fibers are disrupted).
**Demonstrates the reticular fibers (in [[cirrhosis]] the fibers are disrupted).
*Before IHC, reticulin was used to differentiate ''sarcomas'' from ''carcinomas'':<ref name=pmid13536209>{{cite journal |author=MACKENZIE DH |title=Reticulin patterns in the diagnosis of carcinomas and sarcomas |journal=Br. J. Cancer |volume=12 |issue=1 |pages=14–9 |year=1958 |month=March |pmid=13536209 |pmc=2074006 |doi= |url=}}</ref>
*Before IHC, reticulin was used to differentiate ''sarcomas'' from ''carcinomas'':<ref name=pmid13536209>{{cite journal |author=MACKENZIE DH |title=Reticulin patterns in the diagnosis of carcinomas and sarcomas |journal=Br. J. Cancer |volume=12 |issue=1 |pages=14–9 |year=1958 |month=March |pmid=13536209 |pmc=2074006 |doi= |url=}}</ref>
**Sarcomas have reticulin around ''each'' cell.
**Sarcomas have reticulin around ''each'' cell.
**Carcinomas have reticulin around clusters of cells.
**Carcinomas have reticulin around clusters of cells.
*Commonly used in neuropathology.
** In adenoma, reticulin highlights the lost acinar structure of normal pituitary gland.
** Paraganglioma (Zellballen architecture)
** Separating schwannoma (basement membrane around each cell) from meingioma in cerebellopontine angle.
** Separating desmoplastic medulloblastoma from classic/anaplastic forms.


===Interpretation===
===Interpretation===
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Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF]</ref>
Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF]</ref>


==Cresyl violet==
====Images====
<gallery>
Image:Liver_reticulin.jpg | [[Liver]]. Reticulin stain. (WC)
Image:Hepatic_adenoma_high_mag_reticulin.jpg | [[Hepatic adenoma]]. Reticulin stain. (WC)
File:Zellballen paraganglioma.jpg | Reticulin stain highlighting the "Zellballen" architecture of paraganglioma. (WC/jensflorian)
File:Desmoplastic medulloblastoma reticulin stain pale island.jpg | Reticulin staina round the "pale islands" of a desmoplastic medulloblastoma. (WC/jensflorian)
</gallery>
 
==Cresyl violet stain==
===Use===
*Used at some places (e.g. SMH) to look for Helicobacter organisms.
 
===Interpretation===
*Everything is shades of blue.
*Everything is shades of blue.
*Used at some places (e.g. SMH) to look for Helicobacter organisms.
**Helicobacter stains blue.
**Helicobacter stains blue.


==Prussian blue stain==
==Prussian blue stain==
*AKA ''Perl's iron stain''.
*AKA ''Perl's iron stain''.
===Use===
*Useful for iron and hemosiderin; useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
*Useful for iron and hemosiderin; useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
===Interpretation===
*Blue = iron.


Image:
Image:
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*Described well by ''vetmed.vt.edu''.<ref>Prussian blue stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm [http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm]. Accessed on: 5 May 2010.</ref>
*Described well by ''vetmed.vt.edu''.<ref>Prussian blue stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm [http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm]. Accessed on: 5 May 2010.</ref>
*DDx of brown pigment: Fontana-Masson (melanin), Kluver-Barrera stain (lipofuscin).
*DDx of brown pigment: Fontana-Masson (melanin), Kluver-Barrera stain (lipofuscin).
====Images====
<gallery>
Image:Hemosiderosis_high_mag.jpg | Liver [[hemosiderosis]]. Prussian blue stain. (WC/Nephron)
File:Siderophage iron stain CSF.jpg | CSF Siderophages in subarachnoid hemorrhage. (WC/jensflorian)
</gallery>


==Kluver-Barrera stain==
==Kluver-Barrera stain==
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*Useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
*Useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
**Stains lipofuscin.
**Stains lipofuscin.
*Useful to detect demyelinating lesions in the CNS.
Notes:
*[[PAS stain|PAS]] also stains lipofuscin and is more commonly available.


===Interpretation===
===Interpretation===
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*Described well by ''vetmed.vt.edu''.<ref>Kluver-Barrera stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm]. Accessed on: 5 May 2010.</ref>
*Described well by ''vetmed.vt.edu''.<ref>Kluver-Barrera stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm]. Accessed on: 5 May 2010.</ref>
*DDx of brown pigment: Fontana-Masson (melanin), Prussian blue stain (hemosiderin).
*DDx of brown pigment: Fontana-Masson (melanin), Prussian blue stain (hemosiderin).
<gallery>
File:MS Demyelinisation KB 10x.jpg | Encephalomyelitis disseminata (Klüver-Barrera)
</gallery>


==Oil red O stain==
==Oil red O stain==
===Use===
===Use===
*Stain adipose tissue.
*Stains adipose tissue.
*Corroborate diagnosis of [[lipoid pneumonia]].<ref name=pmid25374742>{{Cite journal  | last1 = Yampara Guarachi | first1 = GI. | last2 = Barbosa Moreira | first2 = V. | last3 = Santos Ferreira | first3 = A. | last4 = Sias | first4 = SM. | last5 = Rodrigues | first5 = CC. | last6 = Teixeira | first6 = GH. | title = Lipoid pneumonia in a gas station attendant. | journal = Case Rep Pulmonol | volume = 2014 | issue =  | pages = 358761 | month =  | year = 2014 | doi = 10.1155/2014/358761 | PMID = 25374742 }}</ref>
*Screen for [[GERD]] - positive staining seen in macrophages from [[BAL]] specimens.<ref name=pmid20466562>{{Cite journal  | last1 = Hopkins | first1 = PM. | last2 = Kermeen | first2 = F. | last3 = Duhig | first3 = E. | last4 = Fletcher | first4 = L. | last5 = Gradwell | first5 = J. | last6 = Whitfield | first6 = L. | last7 = Godinez | first7 = C. | last8 = Musk | first8 = M. | last9 = Chambers | first9 = D. | title = Oil red O stain of alveolar macrophages is an effective screening test for gastroesophageal reflux disease in lung transplant recipients. | journal = J Heart Lung Transplant | volume = 29 | issue = 8 | pages = 859-64 | month = Aug | year = 2010 | doi = 10.1016/j.healun.2010.03.015 | PMID = 20466562 }}</ref>
*Uncommon.


Notes:
Notes:
*Must be done on fresh tissue, i.e. it cannot be fixed in formulin.
*Must be done on fresh tissue, i.e. it cannot be fixed in [[formalin]].
 
===Interpretation===
*Red = fat.
 
====Images====
<gallery>
Image:Differentiated_3T3-L1_Cell_line_stained_with_Oil_O_Red.jpg | Oil red O stain. (WC)
</gallery>


==Warthin-Starry stain==
==Warthin-Starry stain==
Line 255: Line 401:
*Background - yellow.
*Background - yellow.


Image:
====Image====
*[http://commons.wikimedia.org/wiki/File:Pylorigastritis.jpg Helicobacter gastritis - Warthin-Starry stain (WC)].
<gallery>
 
Image:Pylorigastritis.jpg | Helicobacter gastritis - Warthin-Starry stain. (WC)
</gallery>
Notes:
Notes:
*Considered a "dirty" stain - picks-up junk in the background.<ref>DB. 4 August 2010.</ref>
*Considered a "dirty" stain - picks-up junk in the background.<ref>DB. 4 August 2010.</ref>
Line 264: Line 411:
Considered a variant of the ''Steiner stain''.<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Overview/80327 http://www.mayomedicallaboratories.com/test-catalog/Overview/80327]. Accessed on: 8 August 2010.</ref>
Considered a variant of the ''Steiner stain''.<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Overview/80327 http://www.mayomedicallaboratories.com/test-catalog/Overview/80327]. Accessed on: 8 August 2010.</ref>
===Use===
===Use===
*Find spirochetes, e.g. syphilis (Treponema pallidum),<ref name=Ref_WMSP455>{{Ref WMSP|455}}</ref> donovan bodies (leishmaniasis),<ref>URL: [http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain]. Accessed on: 4 August 2010.</ref> Helicobacter pylori and Bartonella henselae (Cat-scratch disease).<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Overview/80327 http://www.mayomedicallaboratories.com/test-catalog/Overview/80327]. Accessed on: 8 August 2010.</ref>
*Find spirochetes, e.g. [[syphilis]] (Treponema pallidum),<ref name=Ref_WMSP455>{{Ref WMSP|455}}</ref> donovan bodies (leishmaniasis),<ref>URL: [http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain]. Accessed on: 4 August 2010.</ref> Helicobacter pylori and Bartonella henselae (Cat-scratch disease).<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Overview/80327 http://www.mayomedicallaboratories.com/test-catalog/Overview/80327]. Accessed on: 8 August 2010.</ref>


===Interpretation===
===Interpretation===
Line 270: Line 417:
*Background - yellow.
*Background - yellow.


Images: [http://en.wikipedia.org/wiki/File:Treponema_pallidum_-_very_high_mag_-_extreme_crop.jpg Dieterle stain - T. pallidum (WC)], [http://pathmicro.med.sc.edu/trepo.jpg Treponema (med.sc.edu)], [http://pathmicro.med.sc.edu/fox/spiro-neisseria.htm Spirochetes - several images (med.sc.edu)].
====Images====
<gallery>
Image:Treponema_pallidum_-_very_high_mag_-_extreme_crop.jpg | Dieterle stain - T. pallidum. (WC)
</gallery>
www:
*[http://pathmicro.med.sc.edu/trepo.jpg Treponema (med.sc.edu)].
*[http://pathmicro.med.sc.edu/fox/spiro-neisseria.htm Spirochetes - several images (med.sc.edu)].


==Bielschowsky stain==
==Bielschowsky stain==
Line 283: Line 436:
*Brown/dark brown = plaque, vascular amyloid.
*Brown/dark brown = plaque, vascular amyloid.
*Yellow/brown = other.
*Yellow/brown = other.
Image: [http://commons.wikimedia.org/wiki/File:Cerebellum_-_biel_-_very_high_mag.jpg Bielschowsky stain (WC)].


Ref.: <ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF]</ref>
Ref.: <ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF]</ref>
 
====Image====
==Mucicarmine==
<gallery>
Image:Cerebellum_-_biel_-_very_high_mag.jpg | Bielschowsky stain. (WC/Nephron)
</gallery>
==Mucicarmine stain==
*Stains some mucins... uses the dye ''carmine''.
*Stains some mucins... uses the dye ''carmine''.


Line 301: Line 455:
**Red = mucin.<ref>[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg]</ref>
**Red = mucin.<ref>[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg]</ref>


Images:
====Images====
<gallery>
Image:Cryptococcosis of lung in patient with AIDS. Mucicarmine stain 962 lores.jpg | [[Cryptococcosis]]. Mucicarmine stain. (WC/CDC)
</gallery>
www:
*[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/Pages/page6.htm Mucicarmine stained bowel (medschool.lsuhsc.edu)].  
*[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/Pages/page6.htm Mucicarmine stained bowel (medschool.lsuhsc.edu)].  
*[http://www.nature.com/modpathol/journal/v14/n5/fig_tab/3880332f4.html Mucicarmine stained pancreatic adenosquamous carcinoma (nature.com)].
*[http://www.nature.com/modpathol/journal/v14/n5/fig_tab/3880332f4.html Mucicarmine stained pancreatic adenosquamous carcinoma (nature.com)].


==Alcian blue==
==Alcian blue stain==
===General===
===General===
*Stains acidic mucin (pH=2.5); '''A'''lcian blue = '''A'''cidic.
*Stains acidic mucin (pH=2.5); '''A'''lcian blue = '''A'''cidic.
**A variant uses pH=1.0.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>
**A variant uses pH=1.0.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>


Note:
*''Alcian blue'' (not otherwise specified) usu. refers to the pH=2.5.<ref>URL: [http://www.pathologyoutlines.com/topic/stainsalcianblue.html http://www.pathologyoutlines.com/topic/stainsalcianblue.html]. Accessed on: 11 October 2012.</ref>
===Use===
*Identify ''[[intestinal metaplasia]]'' in the [[intestinal metaplasia of the stomach|stomach]] and [[Barrett esophagus|esophagus]] -- goblets = blue.
Note:
*Esophageal submucosal glands - alcian blue positive.
===Interpretation===
*Blue = acidic mucins.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF]. Accessed on: 20 December 2011.</ref>
Notes:
*Mucin stains:
**[[Alcian blue stain]], [[PASD stain]], [[Mucicarmine stain]].
====Image====
<gallery>
Image:Barrett's_mucosa,_PAS-Alcian_blue_stain.jpg | [[Barrett's esophagus|Barrett's type mucosa]]. Alcian blue stain. (WC/AFIP)
Image:Barrett's_mucosa,_higher_magnification,_Alcian_blue_stain_.jpg | [[Barrett's esophagus|Barrett's type mucosa]]. Alcian blue stain. (WC/AFIP)
</gallery>
==Sodium sulphate-alcian blue stain==
:''Sulfated alcian blue'' (abbreviated '''SAB'') redirects here.
===Use===
===Use===
*Identify ''intestinal metaplasia'' in the [[stomach]] -- goblets = blue.
*Identify [[amyloid]].<ref name=pmid55419>{{Cite journal  | last1 = Pomerance | first1 = A. | last2 = Slavin | first2 = G. | last3 = McWatt | first3 = J. | title = Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology. | journal = J Clin Pathol | volume = 29 | issue = 1 | pages = 22-6 | month = Jan | year = 1976 | doi =  | PMID = 55419 }}</ref><ref>URL: [http://www.polyrnd.com/products/reagent-assembly-kits/conventional/amyloid-stain---sulfated-alcian-blue-(sab).aspx http://www.polyrnd.com/products/reagent-assembly-kits/conventional/amyloid-stain---sulfated-alcian-blue-(sab).aspx]. Accessed on: October 15, 2014.</ref>
*Less [[specificity|specific]] than [[congo red stain|congo red]] but equally [[sensitivity|sensitive]].
*Does not require polarized light.


===Interpretation===
===Interpretation===
*Blue = acidic mucins.
*Green = amyloid.<ref name=pmid55419/>
**Other things that are green: [[mast cell]]s, mucoid degeneration, basophilic myofibre degeneration, califications.
*Yellow = background.
 
Image:
*[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC475937/figure/F1/ SAB (nih.gov)].


Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF]</ref>
==Movat's stain==
==Movat's stain==
===Use===
===Use===
Line 333: Line 521:


How to remember? A.: Primary colours (red, blue, yellow) + black.
How to remember? A.: Primary colours (red, blue, yellow) + black.
==Masson's trichrome==
 
===General use===
====Images====
<gallery>
Image:Cardiac_amyloidosis_very_high_mag_movat.jpg | [[Cardiac amyloidosis]] - Movat stain. (WC/Nephron)
Image:Cystic_medial_degeneration_-_movat_-_low_mag.jpg | [[Cystic medial degeneration]] - Movat stain - low mag. (WC/Nephron)
Image:Cystic_medial_degeneration_-_movat_-_intermed_mag.jpg | Cystic medial degeneration - Movat stain - intermed. mag. (WC/Nephron)
Image:Cystic medial degeneration - movat - high mag.jpg | Cystic medial degeneration - Movat stain - high mag. (WC/Nephron)
</gallery>
 
==Masson's trichrome stain==
*Should '''not''' be confused with the ''[[Mallory trichrome stain]]''.
*May be referred to as ''[[trichrome stain]]''.
===General===
*Collagen vs. muscle.
*Collagen vs. muscle.
===Site===
*Kidney Bx (to assess for fibrosis).
*Liver Bx (to assess for [[cirrhosis]]).


===Interpretation===
===Interpretation===
Line 345: Line 540:
*Red = muscle (smooth muscle actin).
*Red = muscle (smooth muscle actin).
*Baby blue = collagen.
*Baby blue = collagen.
*Blue gray = [[amyloid]].<ref name=pmid33002919>{{cite journal |authors=Kunnath-Velayudhan S, Larsen BT, Coley SM, De Michele S, Santoriello D, Colby TV, Bhagat G, Saqi A |title=Masson Trichrome and Sulfated Alcian Blue Stains Distinguish Light Chain Deposition Disease From Amyloidosis in the Lung |journal=Am J Surg Pathol |volume=45 |issue=3 |pages=405–413 |date=March 2021 |pmid=33002919 |doi=10.1097/PAS.0000000000001593 |url=}}</ref>


Notes: <Ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF]</ref>
Notes: <Ref>ULR: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF]. Accessed on: 2 November 2011.</ref>


===Elastic trichrome===
===Elastic trichrome stain===
General:
General:
*"Elastic trichrome" is one important variant of ''Masson's trichrome''.
*"Elastic trichrome" is one important variant of ''Masson's trichrome''.
Line 355: Line 551:
*Black = nuclei and '''elastin'''.
*Black = nuclei and '''elastin'''.


==HOPS stain==
==Mallory trichome stain==
*The name is an acronym for... ''haematoxylin'', ''orcein'', ''phyloxin'' and ''saffron''.<ref>{{cite journal |author=Perry JR, Bilbao JM, Gray T |title=Fatal basilar vasculopathy complicating bacterial meningitis |journal=Stroke |volume=23 |issue=8 |pages=1175–8 |year=1992 |pmid=1636194 |doi=}} [http://stroke.ahajournals.org/cgi/reprint/23/8/1175.pdf Free Full Text].</ref>
*Should '''not''' be confused with ''[[Masson trichrome stain]]''.
*May be referred to as ''[[trichrome stain]]''.
 
===General===
*Collagen vs. muscle.
*May be done with elastin.
 
===Site===
*Kidney Bx (to assess for fibrosis).
**Considered better than the [[Masson trichrome stain]].
*Liver Bx (to assess for [[cirrhosis]]).
*Cardiovascular/lung (to see differentiate the layers of the arteries, and arteries from veins).
 
===Interpretation===
*Black = nuclei.
*Red = muscle (smooth muscle actin).
*Green = collagen.
 
====Image====
<gallery>
Image:Cirrhosis_high_mag.jpg | [[Cirrhosis]]. Mallory trichrome. (WC/Nephron)
</gallery>
==Haematoxylin orcein phyloxin saffron stain==
*Abbreviated ''HOPS''.<ref name=pmid1636194>{{cite journal |author=Perry JR, Bilbao JM, Gray T |title=Fatal basilar vasculopathy complicating bacterial meningitis |journal=Stroke |volume=23 |issue=8 |pages=1175–8 |year=1992 |pmid=1636194 |doi=}} [http://stroke.ahajournals.org/cgi/reprint/23/8/1175.pdf Free Full Text].</ref>
*It should ''not'' be confused with the ''[[HPS stain]]''.
*It should ''not'' be confused with the ''[[HPS stain]]''.


Line 365: Line 584:
*Yellow (saffron) = collagen.
*Yellow (saffron) = collagen.


==Jones' stain==
==Jones stain==
*AKA ''PAS methenamine technique''.<ref name=pmid>{{Cite journal  | last1 = Jones | first1 = DB. | title = Nephrotic glomerulonephritis. | journal = Am J Pathol | volume = 33 | issue = 2 | pages = 313-29 | month =  | year =  | doi =  | PMID = 13402889 | PMC = 1934622 }}</ref>
*[[AKA]] ''Methenamine PAS'', abbreviated ''MPAS''.
 
===Use===
===Use===
*Visualize basement membrane in kidney biopsies.
*Visualize basement membrane in kidney biopsies.
**Especially useful for the diagnosis of [[membranous nephropathy]] (MN).


===Interpretation===
===Interpretation===
Line 374: Line 597:
*Pink = other structures/background.
*Pink = other structures/background.


Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF]</ref>
Notes:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF]. Accessed on: 19 May 2011.</ref>
 
====Images====
<gallery>
Image:Membranous_nephropathy_-_mpas_-_very_high_mag.jpg | [[Membranous nephropathy|MN]] demonstrated with a MPAS - very high mag. (WC/Nephron)
Image:Membranous_nephropathy_-_cropped_-_mpas_-_very_high_mag.jpg | MN demonstrated with a MPAS - very high mag. (WC/Nephron)
</gallery>


==Hale's colloidal iron==
==Hale's colloidal iron stain==
===Use===
*[[Renal cell carcinoma]] vs. oncocytoma; +ve in chromophobe [[renal cell carcinoma]].<ref name=pmid9537468>{{cite journal |author=Tickoo SK, Amin MB, Zarbo RJ |title=Colloidal iron staining in renal epithelial neoplasms, including chromophobe renal cell carcinoma: emphasis on technique and patterns of staining |journal=Am. J. Surg. Pathol. |volume=22 |issue=4 |pages=419–24 |year=1998 |month=April |pmid=9537468 |doi= |url=http://meta.wkhealth.com/pt/pt-core/template-journal/lwwgateway/media/landingpage.htm?issn=0147-5185&volume=22&issue=4&spage=419}}</ref>


===Interpretation===
{{Main|Hale's colloidal iron stain}}
*Blue is positive.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>


Notes:
==von Kossa stain==
*A few staff think this is a totally useless stain.<ref>ALS several occasions - 2009.</ref>
===General===
*Type of silver stain.<ref name=pmid8360080>{{Cite journal  | last1 = Rungby | first1 = J. | last2 = Kassem | first2 = M. | last3 = Eriksen | first3 = EF. | last4 = Danscher | first4 = G. | title = The von Kossa reaction for calcium deposits: silver lactate staining increases sensitivity and reduces background. | journal = Histochem J | volume = 25 | issue = 6 | pages = 446-51 | month = Jun | year = 1993 | doi =  | PMID = 8360080 }}</ref>


==von Kossa==
===Use===
===Use===
*Look for calcium.
*Look for calcium.
*Actually stains phosphates and carbonates as a surrogate for calcium.


===Interpretation===
===Interpretation===
*Black = calcium.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>
*Black = calcium.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>


==Toluidine blue==
==Toluidine blue stain==
===Use===
===Use===
*May be useful in kidney biopsies.<ref name=pmid16799480>{{cite journal |author=Fischer EG, Moore MJ, Lager DJ |title=Fabry disease: a morphologic study of 11 cases |journal=Mod. Pathol. |volume=19 |issue=10 |pages=1295–301 |year=2006 |month=October |pmid=16799480 |doi=10.1038/modpathol.3800634 |url=http://www.nature.com/modpathol/journal/v19/n10/abs/3800634a.html}}</ref>
*May be useful in kidney biopsies.<ref name=pmid16799480>{{cite journal |author=Fischer EG, Moore MJ, Lager DJ |title=Fabry disease: a morphologic study of 11 cases |journal=Mod. Pathol. |volume=19 |issue=10 |pages=1295–301 |year=2006 |month=October |pmid=16799480 |doi=10.1038/modpathol.3800634 |url=http://www.nature.com/modpathol/journal/v19/n10/abs/3800634a.html}}</ref><ref name=pmid21659728>{{Cite journal  | last1 = Nicholas | first1 = SB. | last2 = Basgen | first2 = JM. | last3 = Sinha | first3 = S. | title = Using stereologic techniques for podocyte counting in the mouse: shifting the paradigm. | journal = Am J Nephrol | volume = 33 Suppl 1 | issue =  | pages = 1-7 | month =  | year = 2011 | doi = 10.1159/000327564 | PMID = 21659728 }}</ref>
*Stains [[mast cell]]s, [[pneumocystis jirovecii]].
*Stains [[mast cell]]s, [[pneumocystis jirovecii]].


===Interpretation===
===Interpretation===
*Dark blue - nuclei, mast cell granules.
*Dark blue - nuclei, mast cell granules (darker than nuclei).
*Light blue - cytoplasm.
*Light blue - cytoplasm.
*Red/magneta - cartilage. (???)


Ref.: looks a bit sketchy - <ref>[http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html]</ref>
Refs: looks a bit sketchy<ref>URL: [http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html]. Accessed on: 17 March 2011.</ref>, <ref>URL: [http://www.dermnetnz.org/doctors/dermatopathology/stains.html http://www.dermnetnz.org/doctors/dermatopathology/stains.html]. Accessed on: 17 March 2011.</ref>


Image:
====Image====
*[http://commons.wikimedia.org/wiki/File:Smear_of_Pneumocystis_carinii._Toluidine_blue_stain_PHIL_596_lores.jpg PCP stained with toluidine blue (mediawiki.org)].
<gallery>
Image:Smear_of_Pneumocystis_carinii._Toluidine_blue_stain_PHIL_596_lores.jpg | [[PCP]] stained with toluidine blue. (WC)
</gallery>
www:
*[http://www.biomedcentral.com/1471-2407/5/121/figure/F3?highres=y Mast cells stained with toluidine blue (biomedcentral.com)].
*[http://www.biomedcentral.com/1471-2407/5/121/figure/F3?highres=y Mast cells stained with toluidine blue (biomedcentral.com)].


Line 412: Line 643:
*Many variants of this stain exist.
*Many variants of this stain exist.
*Specimens are air-dried.
*Specimens are air-dried.


Interpretation:<ref>{{cite journal |author=Horobin RW, Walter KJ |title=Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears |journal=Histochemistry |volume=86 |issue=3 |pages=331–6 |year=1987 |pmid=2437082 |doi= |url=http://www.springerlink.com/content/r81x25451m841866/}}</ref>
Interpretation:<ref>{{cite journal |author=Horobin RW, Walter KJ |title=Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears |journal=Histochemistry |volume=86 |issue=3 |pages=331–6 |year=1987 |pmid=2437082 |doi= |url=http://www.springerlink.com/content/r81x25451m841866/}}</ref>
Line 419: Line 649:
*Purple - nuclear chromatin, neutrophil granules, platelets.
*Purple - nuclear chromatin, neutrophil granules, platelets.


===Field's stain===
===Field stain===
*Variant of the ''Romanowsky stain'' for rapid processing.
*Variant of the ''Romanowsky stain'' for rapid processing.
*Tends to "blow-up" cell, i.e. cells are larger vis-a-vis ''[[Pap stain]]''.
*Tends to "blow-up" cell, i.e. cells are larger vis-a-vis ''[[Pap stain]]''.


===Diff-Quik===
===Diff-Quik===
:Pronounced ''Diff-Quick''.
*Proprietary variant of ''Romanowsky stain''.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm]. Accessed on: 4 January 2010.</ref>
*Proprietary variant of ''Romanowsky stain''.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm]. Accessed on: 4 January 2010.</ref>
*Use: cytopathology.
 
Uses:
*[[Cytopathology]].
*[[Helicobacter gastritis]] - organisms are dark blue against a light blue background.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm]. Accessed on: 30 August 2012.</ref>
 
====Images====
<gallery>
Image: Lung adenocarcinoma - Diff-Quik -- high mag.jpg | [[Pulmonary_cytopathology#Adenocarcinoma|Lung adenocarcinoma]] - DQ - high mag. (WC)
Image: Lung adenocarcinoma - Diff-Quik -- very high mag.jpg | Lung adenocarcinoma - DQ - very high mag. (WC)
Image: Lung small cell carcinoma - Diff-Quik -- very high mag.jpg | Lung SmCC - DQ - very high mag. (WC)
Image: Lung small cell carcinoma - Diff-Quik -- extremely high mag.jpg | Lung SmCC - DQ - extremely high mag. (WC)
</gallery>


===Wright stain===
===Wright stain===
Line 433: Line 675:
*Blood films.
*Blood films.


===May-Grünwald-Giemsa===
===May-Grünwald-Giemsa stain===
*A variant of the ''Romanowsky stain''; popular in Europe.
*A variant of the ''Romanowsky stain''; popular in Europe.
*Abbreviated ''MGG''.
*Abbreviated ''MGG''.
Line 442: Line 684:


==Papanicolaou stain==
==Papanicolaou stain==
*Often abbreviated ''Pap stain''.
*Abbreviated ''Pap stain''.
*Can be thought of as the H&E of cytopathology.
{{Main|Papanicolaou stain}}
**It is a modified H&E stain.
*Specimens are fixed in ethanol.
*Good for seeing nuclear detail.
*Out-of-focus cytoplasm is translucent; allows one to focus overlapped cells in different planes.
 
===Use===
*Cytopathology.
 
===Interpretation===
*Blue/purple = nucleus.
*Green/pink = cytoplasm.
*Orange = keratin.
 
Image:
*[http://commons.wikimedia.org/wiki/File:Urine_citology_urothelial_carcinoma_2.jpg Pap stain - urine cytology (wikimedia.org)].
 
==Fontana-Masson==
*AKA ''Fontana-Masson stain for melanin'', ''melanin stain''.
*A type of silver stain.
 
Stains:
#Melanin.
#"argentaffin granules" of the digestive tract.
 
===Use===
*Stain for melanin.
**Used to differentiate brown pigments (lipofuscin, hemosiderin, melanin).<ref>URL: [http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm]. Accessed on: 5 May 2010.</ref>
***Many pathologists prefer IHC, i.e. ''Melan A'' over this stain.


Image:
==Fontana-Masson stain==
*[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm Fontana-Masson stain (vetmed.vt.edu)].
*[[AKA]] ''Masson-Fontana stain'',<ref name=pmid16081962>{{Cite journal  | last1 = Gaitanis | first1 = G. | last2 = Chasapi | first2 = V. | last3 = Velegraki | first3 = A. | title = Novel application of the masson-fontana stain for demonstrating Malassezia species melanin-like pigment production in vitro and in clinical specimens. | journal = J Clin Microbiol | volume = 43 | issue = 8 | pages = 4147-51 | month = Aug | year = 2005 | doi = 10.1128/JCM.43.8.4147-4151.2005 | PMID = 16081962 }}</ref> ''Fontana-Masson stain for melanin'', ''melanin stain''.
{{Main|Fontana-Masson stain}}


==Schmorl's stain==
==Schmorl's stain==
Line 482: Line 697:
Notes:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: 5 May 2010.</ref>
Notes:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: 5 May 2010.</ref>


==Martius scarlet blue==
==Martius scarlet blue stain==
===General===
===General===
*Stains connective tissue and fibrin.<ref>URL: [http://www.bris.ac.uk/vetpath/cpl/msb.html http://www.bris.ac.uk/vetpath/cpl/msb.html]. Accessed on: 26 November 2010.</ref>
*Stains connective tissue and fibrin.<ref>URL: [http://www.bris.ac.uk/vetpath/cpl/msb.html http://www.bris.ac.uk/vetpath/cpl/msb.html]. Accessed on: 26 November 2010.</ref>
Line 496: Line 711:


Image:
Image:
*[http://www.bris.ac.uk/vetpath/cpl/msb.jpg MSB (bris.ac.uk)].
*[https://www.ole.bris.ac.uk/bbcswebdav/institution/Faculty%20of%20Health%20Sciences/Veterinary%20Science/eLearning%20resources/Pathology%20Laboratory%20Protocols/hst/msb.html MSB (bris.ac.uk)].


Ref.:<ref>URL: [http://www.bris.ac.uk/vetpath/cpl/msb.html http://www.bris.ac.uk/vetpath/cpl/msb.html]. Accessed on: 26 November 2010.</ref>
Ref.:<ref>URL: [http://www.bris.ac.uk/vetpath/cpl/msb.html http://www.bris.ac.uk/vetpath/cpl/msb.html]. Accessed on: 26 November 2010.</ref>


==Picro-Mallory==
==Picro-Mallory stain==
===General===
===General===
*Find fibrin.
*Find fibrin.
Line 512: Line 727:
*[http://4.bp.blogspot.com/_rlWIjG4vffU/S3FC7BNIJHI/AAAAAAAACDc/XfOz9X_bvbY/s1600-h/publicado+lengua%233.jpg Picro-Mallary (blogspot.com)].
*[http://4.bp.blogspot.com/_rlWIjG4vffU/S3FC7BNIJHI/AAAAAAAACDc/XfOz9X_bvbY/s1600-h/publicado+lengua%233.jpg Picro-Mallary (blogspot.com)].


==Verhoeff-van Gieson Stain==
==Verhoeff-van Gieson stain==
:''Verhoeff stain'' redirects here.
*[[AKA]] Elastic van Gieson stain, abbreviated ''EVG''.
===General===
===General===
*[[AKA]] Elastic van Gieson stain.
**Abbreviated ''EVG''.
*Similar to ''Masson Trichrome & Verhoeff stain''.<ref>URL: [http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm]. Accessed on: 3 January 2011.</ref>
*Similar to ''Masson Trichrome & Verhoeff stain''.<ref>URL: [http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm]. Accessed on: 3 January 2011.</ref>


Line 525: Line 740:
*Collagen = bright red.
*Collagen = bright red.
*Muscle = dull red.
*Muscle = dull red.
<gallery>
File:Cerebral aneurysm EVG stain.jpg |EVG stain of a cerebral aneurysm. (WC/jensflorian)
</gallery>


==Copper stain==
==Copper stain==
===General===
===General===
*Used in liver biopsies.
*Used in liver biopsies.
*May be seen in Wilson's disease.
*May be seen in [[Wilson's disease]].


Note:
Note:
Line 541: Line 760:


==Shikata stain==
==Shikata stain==
*[[AKA]] Orcein Stain for Copper-Protein, [[AKA]] Shikata-Cu,<ref>[http://www.mayomedicallaboratories.com/test-catalog/Overview/9836 http://www.mayomedicallaboratories.com/test-catalog/Overview/9836]. Accessed on: 24 January 2011.</ref> [[AKA]] Shikata's orcein staining.<ref>URL: [http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat]. Accessed on: 24 January 2011.</ref>
*[[AKA]] Orcein stain for copper-protein.
*[[AKA]] Shikata-Cu,<ref>[http://www.mayomedicallaboratories.com/test-catalog/Overview/9836 http://www.mayomedicallaboratories.com/test-catalog/Overview/9836]. Accessed on: 24 January 2011.</ref>  
*[[AKA]] Shikata's orcein staining.<ref>URL: [http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat]. Accessed on: 24 January 2011.</ref>


===General===
===General===
*Used in [[medical liver disease|medical liver]] biopsies - used to find copper.  
*Used in [[medical liver disease|medical liver]] biopsies - stains sulfhydrl groups and identifies:
**Copper-associated protein.
**Elastin.
**[[Hepatitis B]] surface antigen.<ref name=pmid7822848>{{Cite journal  | last1 = Ghosh | first1 = AK. | last2 = Dasgupta | first2 = A. | last3 = Raha | first3 = K. | last4 = Jana | first4 = A. | last5 = Majumdar | first5 = DN. | title = Hepatic histology in chronic liver disease in hepatitis B surface antigen positive cases. | journal = J Indian Med Assoc | volume = 92 | issue = 10 | pages = 333-5 | month = Oct | year = 1994 | doi =  | PMID = 7822848 }}</ref>


===Interpretation===
===Interpretation===
Features:<ref>URL: [http://www.nottingham.ac.uk/pathology/protocols/shikata.html http://www.nottingham.ac.uk/pathology/protocols/shikata.html]. Accessed on: 24 January 2011.</ref>
Features:<ref>URL: [http://www.nottingham.ac.uk/pathology/protocols/shikata.html http://www.nottingham.ac.uk/pathology/protocols/shikata.html]. Accessed on: 24 January 2011.</ref>
*Purple/brown = elastin fibres.
*Dark purple/brown = elastin fibres, HBsAg and copper-associated protein
*Red = Copper associated protein.
*Light purple = background
*Red = nuclei (only if counter-stain used)
 
==Gömöri Trichrome stain==
*Named after George Gömöri<ref>GOMORI, G. - A rapid one-step trichrome stain. Am. J. Clin. Path. 20: 661-664, 1950.</ref>
 
===General===
*Used in [[muscle biopsies]] - used to find abnormal mitochondrial deposits.
 
===Interpretation===
*Dark green = muscle fibers.
*Red = nuclei.
*Bright red = mitochondria, red blood cells.
 
Images:
<gallery>
File:Ragged red fibers in MELAS.jpg | Ragged red fibers in MELAS, a mitochondrial disease. (WC)
File:Dilated peri-tubular capillaries filled with sickled RBCs, original Gomori's trichrome stain.jpg | Sickle cell nephropathy. (WC)
</gallery>
 
==Miller stain==
===General===
*Stains elastin.
*Used to identify blood vessels and [[pleural invasion]] in [[lung cancer]].
 
===Interpretation===
Staining:<ref>URL: [https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm]. Accessed on: 28 August 2015.</ref>
*Black = elastin fibres, granules in mast cells.
*Red = collagen.
*Yellow = muscle, fibrin, [[erythrocytes]].
*Green/brown = nuclei.
 
====Images====
<gallery>
Image: Normal visceral pleura of lung - Miller -- high mag.jpg | Miller stain showing lung tissue. (WC)
</gallery>


==See also==
=See also=
*[[Immunohistochemistry]].
*[[Immunohistochemistry]].
*[[Basics]].
*[[Basics]].


==References==
=References=
{{reflist|2}}
{{reflist|2}}


==External links==
=External links=
*[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MANUALS.html Procedure manuals] - med.utah.edu.
*[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MANUALS.html Procedure manuals] - med.utah.edu.
*[http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html Special stains (introduction)] - med.utah.edu.
*[http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html Special stains (introduction)] - med.utah.edu.
*[http://www.histology-world.com/stains/stains.htm Stains] - histology-world.com.
*[http://www.histology-world.com/stains/stains.htm Stains] - histology-world.com.


[[Category:Basics of pathology]]
[[Category:Basics]]
[[Category:Stains|Stains]]

Latest revision as of 16:05, 1 May 2024

This article deals with stains. The H&E stain is only one of many stains that are used in pathology.

Non-H&E stains are often referred to as special stains.

Where to start...

Principles

When considering additional (i.e. special) stains one should (in order) do the following:[1]

  1. Make sure one has exhausted the clinical history; history is considered the best special stain.
  2. Special stains (below).
  3. Immunohistochemistry (dealt with in a separate article).
  4. Molecular testing, electron microscopy.

Common stains

  1. H&E stain.
  2. PAS stain.
  3. PAS-D stain.
  4. AFB stains, e.g. Ziehl-Neelsen stain.
  5. Congo red.
  6. GMS stain.
  7. Gram stain.

Immunohistochemistry

General

  • Abbreviated IHC.

Interpretation

Simple version:

  • Positive is (usually): brown.
  • Negative tissue is: light blue.

Important notes:

  • One has to know where the target (of the antibody) is supposed to be, i.e. cytoplasm vs. cell membrane.
  • The edge of the tissue may have light staining - edge effect.
  • If everything is brown... suspect that it didn't work.
  • In some situations you're blessed with an internal control, e.g. in renal tumours CD10 will stain RCC and the proximal tubule, in GISTs - CD117 the mast cells are positive.

Work-up of infection

It often not possible to be definitive by staining.[2]

Basic panel:

  • Gram stain - for bacteria.
  • GMS stain - fungal stain.
  • PAS (or PAS-D) - fungal stain.

Fungi

Fungi are a type of microorganisms. They are seen by pathologist every once in a while.

Specific stains

What follows is a big list... of stains.

Haematoxylin and eosin stain

  • Abbreviated H&E.

Haematoxylin phyloxin saffron stain

General

  • Abbreviated HPS.
  • An alternative to the H&E stain - some pathol. departments use this as their standard.

Interpretation

  • Haematoxylin = blue -- stains nucleus.
  • Phyloxin = pink -- stains muscle and cytoplasm.
  • Saffron = yellow -- stains collagen.
  • An alternative to H&E stain.
    • Fibrosis is easier to see on HPS than H&E... as one can see the collagen.

Images

Periodic acid Schiff stain

  • Abbreviated PAS.

Primary application

  • Kidney biopsies, medical.
  • Liver biopsies, medical.
    • Positive in alpha-1 antitrypsin deficiency.

Utility

  • Stains - lipofuscin,[3] basement membranes, fungi, glycogen, (neutral) mucin.

Interpretation

  • Magenta = glycogen, mucin, fungi.
  • Blue = nuclei.

Ref.:[4]

Image

Periodic acid Schiff fungal stain

  • Abbreviated PASF.

Primary application

Interpretation

  • Light purple = fungi.
  • Light green = background.
  • Washed-out light purple = Gram positive bacilli.

Note:

  • This is much improved over the PAS in the context of skin, as the background is similar to the fungal organisms.

Periodic acid Schiff with diastase

  • Abbreviated: PAS-D and PASD.

General

  • Diastase digests the glycogen.
  • "PAS diastase resistant"[5] implies PASD +ve and PAS +ve.

Use

  • Stains mucin.
  • Used to identify glycogen (together with PAS stain).
    • Glycogen = clear (digested) on PAS-D.
    • Glycogen = magenta on PAS.

Notes: [6]

Interpretation

  • Light purple = fungi.
  • Light blue/pink = background. ???

Gomori methenamine-silver stain

  • Abbreviated GMS.

Note:

  • GMS is "Grocott's methenamine Silver" according to WMSP.[7]

Use

Image

Acid-fast bacilli stains

  • Abbreviated: AFB.

There are several AFB stains:

Ziehl-Neelsen stain

  • Most popular acid-fast bacilli stain.
  • Stains other mycobacteria -- not specific for tuberculosis.
    • Stains Nocardia.[10]

Image

Fite stain

Interpretation:

  • Red = AFB.
  • Blue = background.

Auramine-rhodamine stain

  • Fluorescent stain.

Image

Kinyoun stain

Congo red stain

Use

  • Used to look for amyloid.
    • Mnemonic: CRAP = congo red amyloid protein.
  • An alternate stain for amyloid is Thioflavin T.

Note:

  • Thick sections (~10 micrometers) are considered a requirement for the stain to work properly.[13]
    • If the section is too thin... it doesn't work.

Interpretation

  • Amyloid = pink/red.
  • Nuclei = blue.

Ref.:[14]

Image

Thioflavin T stain

Use

Interpretation

  • Amyloid = green.

Image: Amyloid (inano.au.dk).

Gram stain

Use

  • "It is useless for finding bacteria."[15]
    • If they are to be seen... they'll be visible on H&E.

Note:

  • Microbiology is better at finding organisms than pathology.
    • They have one significant advantage -- if a small amount of bugs are present... they grows into a large (obviously visible) colony.

DDx for common patterns

A short list of bacteria and their characteristics:[16]

Shape\Gram stain Positive Negative Variable or negative
Bacilli Clostridium difficile, Bacillus anthracis, Nocardia spp. Escherichia coli, Helicobacter pylori, Yersinia pestis, Hemophilus influenzae Mycobacterium tuberulosis, Legionella pneumophila[17]
Cocci Streptococcus pneumoniae, Staphylococcus aureus Neisseria meningitidis, Moraxella catarrhalis

Interpretation

  • Purple (or blue) = Gram positive organisms.
  • Red = Gram negative organisms, nuclei.[18]
  • Yellow = background.

Notes:

  • Many of the bacteria are quite small relative to lymphocytes; Escherichia coli is 1-2 micrometers long x 0.25 micrometers in diameter.[19]
  • Epithelial cell nuclei & stromal cell nuclei may stain red.
  • Memory device: purple = positive.

Images

Luxol fast blue stain

  • Abbreviated LFB.

Use

  • Neuropathology, myelin stain.

Intepretation

  • Blue = myelinated fibers (contain lipoproteins), lipofuscin.[20]
    • Lack of blue (where it ought to be) = demyelination.
  • Purple = nerve cell (e.g. neuron).
  • Neutrophils = pink.

Ref.:[21]

Image

Giemsa stain

Use

Interpretation

  • Tissue is light blue/green.
  • Goblet cells are purple.[23]

Image:

Reticulin stain

Use

  • Liver biopsy, medical.
    • Demonstrates the reticular fibers (in cirrhosis the fibers are disrupted).
  • Before IHC, reticulin was used to differentiate sarcomas from carcinomas:[25]
    • Sarcomas have reticulin around each cell.
    • Carcinomas have reticulin around clusters of cells.
  • Commonly used in neuropathology.
    • In adenoma, reticulin highlights the lost acinar structure of normal pituitary gland.
    • Paraganglioma (Zellballen architecture)
    • Separating schwannoma (basement membrane around each cell) from meingioma in cerebellopontine angle.
    • Separating desmoplastic medulloblastoma from classic/anaplastic forms.


Interpretation

  • Black = reticular fibers.
  • Red = nuclei.

Notes:[26]

Images

Cresyl violet stain

Use

  • Used at some places (e.g. SMH) to look for Helicobacter organisms.

Interpretation

  • Everything is shades of blue.
    • Helicobacter stains blue.

Prussian blue stain

  • AKA Perl's iron stain.

Use

  • Useful for iron and hemosiderin; useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).

Interpretation

  • Blue = iron.

Image:

Notes:

  • Described well by vetmed.vt.edu.[27]
  • DDx of brown pigment: Fontana-Masson (melanin), Kluver-Barrera stain (lipofuscin).

Images

Kluver-Barrera stain

Combination of:

  • Luxol Fast Blue,
  • Cresyl Violet,
  • Special component for lipofuscin.

Use

  • Useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
    • Stains lipofuscin.
  • Useful to detect demyelinating lesions in the CNS.

Notes:

  • PAS also stains lipofuscin and is more commonly available.

Interpretation

  • Blue pigmented granules = lipofuscin.

Notes:

  • Described well by vetmed.vt.edu.[28]
  • DDx of brown pigment: Fontana-Masson (melanin), Prussian blue stain (hemosiderin).

Oil red O stain

Use

  • Stains adipose tissue.
  • Corroborate diagnosis of lipoid pneumonia.[29]
  • Screen for GERD - positive staining seen in macrophages from BAL specimens.[30]
  • Uncommon.

Notes:

  • Must be done on fresh tissue, i.e. it cannot be fixed in formalin.

Interpretation

  • Red = fat.

Images

Warthin-Starry stain

Background:

  • Developed by a bunch of pathologists in Michigan to look for spirochetes.[31]

Use

  • Find spirochetes, e.g. syphilis (Treponema pallidum),[32] cat-scratch disease (Bartonella henselae).
  • Find Helicobacter spp., e.g. Helicobacter pylori -- Mount Sinai Hospital.[33]

Interpretation:[34]

  • Spirochetes - black.
  • Background - yellow.

Image

Notes:

  • Considered a "dirty" stain - picks-up junk in the background.[35]

Dieterle stain

Considered a variant of the Steiner stain.[36]

Use

  • Find spirochetes, e.g. syphilis (Treponema pallidum),[37] donovan bodies (leishmaniasis),[38] Helicobacter pylori and Bartonella henselae (Cat-scratch disease).[39]

Interpretation

  • Spirochetes - black.
  • Background - yellow.

Images

www:

Bielschowsky stain

Abbreviated: Biel stain.

Use

  • Stains glial tissue, i.e. brain.
    • Demonstrates neurofibrillary tangles, senile plaques (as in Alzheimer's disease).

Interpretation

  • Black = axons, tangles, plaques.
  • Brown/dark brown = plaque, vascular amyloid.
  • Yellow/brown = other.

Ref.: [40]

Image

Mucicarmine stain

  • Stains some mucins... uses the dye carmine.

Use

  • Identify mucin.
  • Malignant cells that produce mucin... carcinomas.[41]

Interpretation

  • Carmine with metanil yellow and Weigert's Hematoxylin:[42]
    • Blue/black = nucleus.
    • Yellow = background.
    • Red = mucin.[43]

Images

www:

Alcian blue stain

General

  • Stains acidic mucin (pH=2.5); Alcian blue = Acidic.
    • A variant uses pH=1.0.[7]

Note:

  • Alcian blue (not otherwise specified) usu. refers to the pH=2.5.[44]

Use

Note:

  • Esophageal submucosal glands - alcian blue positive.

Interpretation

  • Blue = acidic mucins.[45]

Notes:

Image

Sodium sulphate-alcian blue stain

Sulfated alcian blue (abbreviated 'SAB) redirects here.

Use

Interpretation

  • Green = amyloid.[46]
    • Other things that are green: mast cells, mucoid degeneration, basophilic myofibre degeneration, califications.
  • Yellow = background.

Image:

Movat's stain

Use

Components

  • Acid fuchsin, alcian blue, crocein scarlet, elastic hematoxylin, and saffron.[48][49]

Interpretation of Movat stain

  • Black = nuclei and elastic fibers.
  • Yellow = collagen and reticular fibers.
  • Blue = mucin, ground substance.
  • Red (intense) = fibrin.
  • Red = muscle.

Reference: [49]

How to remember? A.: Primary colours (red, blue, yellow) + black.

Images

Masson's trichrome stain

General

  • Collagen vs. muscle.

Interpretation

  • Black = nuclei.
  • Red = muscle (smooth muscle actin).
  • Baby blue = collagen.
  • Blue gray = amyloid.[50]

Notes: [51]

Elastic trichrome stain

General:

  • "Elastic trichrome" is one important variant of Masson's trichrome.

Interpretation - as above in Masson's trichrome - plus:

  • Black = nuclei and elastin.

Mallory trichome stain

General

  • Collagen vs. muscle.
  • May be done with elastin.

Site

  • Kidney Bx (to assess for fibrosis).
  • Liver Bx (to assess for cirrhosis).
  • Cardiovascular/lung (to see differentiate the layers of the arteries, and arteries from veins).

Interpretation

  • Black = nuclei.
  • Red = muscle (smooth muscle actin).
  • Green = collagen.

Image

Haematoxylin orcein phyloxin saffron stain

  • Abbreviated HOPS.[52]
  • It should not be confused with the HPS stain.

Interpretation

  • Blue (haematoxylin) = nuclei.
  • Black (orcein) = elastin.
  • Red (phyloxin) = muscle.
  • Yellow (saffron) = collagen.

Jones stain

  • AKA PAS methenamine technique.[53]
  • AKA Methenamine PAS, abbreviated MPAS.

Use

  • Visualize basement membrane in kidney biopsies.

Interpretation

  • Black = basement membrane.
  • Blue = nuclei.
  • Pink = other structures/background.

Notes:[54]

Images

Hale's colloidal iron stain

von Kossa stain

General

  • Type of silver stain.[55]

Use

  • Look for calcium.
  • Actually stains phosphates and carbonates as a surrogate for calcium.

Interpretation

  • Black = calcium.[7]

Toluidine blue stain

Use

Interpretation

  • Dark blue - nuclei, mast cell granules (darker than nuclei).
  • Light blue - cytoplasm.
  • Red/magneta - cartilage. (???)

Refs: looks a bit sketchy[58], [59]

Image

www:

Romanowsky stain

  • Occasionally spelled Romanowski.
  • Many variants of this stain exist.
  • Specimens are air-dried.

Interpretation:[60]

  • Red - RBCs, eosinophil granules.
  • Blue (basophilic) - lymphocyte cytoplasm.
  • Purple - nuclear chromatin, neutrophil granules, platelets.

Field stain

  • Variant of the Romanowsky stain for rapid processing.
  • Tends to "blow-up" cell, i.e. cells are larger vis-a-vis Pap stain.

Diff-Quik

Pronounced Diff-Quick.
  • Proprietary variant of Romanowsky stain.[61]

Uses:

Images

Wright stain

  • A variant of the Romanowsky stain; popular in North American.

Use:

  • Blood films.

May-Grünwald-Giemsa stain

  • A variant of the Romanowsky stain; popular in Europe.
  • Abbreviated MGG.

Use:

  • Blood films.
  • Cytopathology.

Papanicolaou stain

  • Abbreviated Pap stain.

Fontana-Masson stain

  • AKA Masson-Fontana stain,[63] Fontana-Masson stain for melanin, melanin stain.

Schmorl's stain

  • Stains melanin.
    • Similar to Fontana-Masson stain.

Notes:[64]

Martius scarlet blue stain

General

  • Stains connective tissue and fibrin.[65]
  • Abbreviated MSB.

Use:

Interpretation

  • Muscle and fibrin - red.
  • Nuclei = brown/black.
  • Collagen - blue.
  • Red blood cells - yellow.

Image:

Ref.:[66]

Picro-Mallory stain

General

  • Find fibrin.

Interpretation[67]

  • Fibrin = red.
  • Erythrocytes = yellow.
  • Connective tissue = blue.

Image:

Verhoeff-van Gieson stain

Verhoeff stain redirects here.
  • AKA Elastic van Gieson stain, abbreviated EVG.

General

  • Similar to Masson Trichrome & Verhoeff stain.[68]

Use:

  • Examine large blood vessels.[69]

Interpretation

  • Elastin = black.
  • Collagen = bright red.
  • Muscle = dull red.

Copper stain

General

Note:

  • Copper staining is a non-specific finding seen in many liver diseases; it is associated with impaired bile secretion.[70]

Interpretation

  • Copper = red granules.

Images:

Shikata stain

  • AKA Orcein stain for copper-protein.
  • AKA Shikata-Cu,[72]
  • AKA Shikata's orcein staining.[73]

General

  • Used in medical liver biopsies - stains sulfhydrl groups and identifies:

Interpretation

Features:[75]

  • Dark purple/brown = elastin fibres, HBsAg and copper-associated protein
  • Light purple = background
  • Red = nuclei (only if counter-stain used)

Gömöri Trichrome stain

  • Named after George Gömöri[76]

General

Interpretation

  • Dark green = muscle fibers.
  • Red = nuclei.
  • Bright red = mitochondria, red blood cells.

Images:

Miller stain

General

Interpretation

Staining:[77]

  • Black = elastin fibres, granules in mast cells.
  • Red = collagen.
  • Yellow = muscle, fibrin, erythrocytes.
  • Green/brown = nuclei.

Images

See also

References

  1. LAE. 13 July 2010.
  2. Woods GL, Walker DH (July 1996). "Detection of infection or infectious agents by use of cytologic and histologic stains". Clin. Microbiol. Rev. 9 (3): 382-404. PMC 172900. PMID 8809467. http://cmr.asm.org/cgi/pmidlookup?view=long&pmid=8809467.
  3. Kovi J, Leifer C (July 1970). "Lipofuscin pigment accumulation in spontaneous mammary carcinoma of A/Jax mouse". J Natl Med Assoc 62 (4): 287–90. PMC 2611776. PMID 5463681. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2611776/pdf/jnma00512-0077.pdf.
  4. http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF
  5. Qizilbash, A.; Young-Pong, O. (Jun 1983). "Alpha 1 antitrypsin liver disease differential diagnosis of PAS-positive, diastase-resistant globules in liver cells.". Am J Clin Pathol 79 (6): 697-702. PMID 6189389.
  6. http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF
  7. 7.0 7.1 7.2 Humphrey, Peter A; Dehner, Louis P; Pfeifer, John D (2008). The Washington Manual of Surgical Pathology (1st ed.). Lippincott Williams & Wilkins. pp. 682. ISBN 978-0781765275.
  8. URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF. Accessed on: 19 May 2011.
  9. Ilyas S, Youssef D, Chaudhary H, Al-Abbadi MA (September 2011). "Myocbacterium-avium intracellulare associated inflammatory pseudotumor of the anterior nasal cavity". Head Neck Pathol 5 (3): 296–301. doi:10.1007/s12105-011-0248-0. PMC 3173547. PMID 21327589. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3173547/.
  10. URL: http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html. Accessed on: 19 May 2011.
  11. Kehl, KS.; Cicirello, H.; Havens, PL. (Feb 1995). "Comparison of four different methods for detection of Cryptosporidium species.". J Clin Microbiol 33 (2): 416-8. PMID 7536216.
  12. Ignatius, R.; Lehmann, M.; Miksits, K.; Regnath, T.; Arvand, M.; Engelmann, E.; Futh, U.; Hahn, H. et al. (Feb 1997). "A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens.". J Clin Microbiol 35 (2): 446-9. PMID 9003613.
  13. URL: http://www.ihcworld.com/_protocols/special_stains/congo_red_bennhold.htm. Accessed on: 26 January 2012.
  14. URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF. Accessed on: 4 December 2010.
  15. St. Michael's Hospital - Stains Handout.
  16. URL: http://www.atsu.edu/faculty/chamberlain/Website/pnebact.htm. Accessed on: 7 May 2013.
  17. URL: http://meded.ucsd.edu/isp/1999/CAP/legion.html. Accessed on: 7 May 2013.
  18. URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF. Accessed on: 7 May 2013.
  19. URL: http://www.lpi.usra.edu/publications/slidesets/marslife/slide_27.html.
  20. MUN. 26 November 2010.
  21. http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF
  22. URL: http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html. Accessed on: April 6, 2009.
  23. URL: http://www.kennedy.ox.ac.uk/facilities/histology/histology-information. Accessed on: 17 August 2015.
  24. URL: http://amser.org/index.php?P=AMSER--ResourceFrame&resourceId=6018. Accessed on: 17 August 2015.
  25. MACKENZIE DH (March 1958). "Reticulin patterns in the diagnosis of carcinomas and sarcomas". Br. J. Cancer 12 (1): 14–9. PMC 2074006. PMID 13536209. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2074006/.
  26. http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF
  27. Prussian blue stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm. Accessed on: 5 May 2010.
  28. Kluver-Barrera stain. URL:http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm. Accessed on: 5 May 2010.
  29. Yampara Guarachi, GI.; Barbosa Moreira, V.; Santos Ferreira, A.; Sias, SM.; Rodrigues, CC.; Teixeira, GH. (2014). "Lipoid pneumonia in a gas station attendant.". Case Rep Pulmonol 2014: 358761. doi:10.1155/2014/358761. PMID 25374742.
  30. Hopkins, PM.; Kermeen, F.; Duhig, E.; Fletcher, L.; Gradwell, J.; Whitfield, L.; Godinez, C.; Musk, M. et al. (Aug 2010). "Oil red O stain of alveolar macrophages is an effective screening test for gastroesophageal reflux disease in lung transplant recipients.". J Heart Lung Transplant 29 (8): 859-64. doi:10.1016/j.healun.2010.03.015. PMID 20466562.
  31. URL: http://www.merriam-webster.com/medical/warthin. Accessed on: 17 August 2010.
  32. URL: http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html. Accessed on: April 6, 2009.
  33. http://www.dako.co.uk/index/prod_search/prod_products.htm?productareaid=41&baseprodidver=A224462007
  34. http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAIN029.html
  35. DB. 4 August 2010.
  36. URL: http://www.mayomedicallaboratories.com/test-catalog/Overview/80327. Accessed on: 8 August 2010.
  37. Humphrey, Peter A; Dehner, Louis P; Pfeifer, John D (2008). The Washington Manual of Surgical Pathology (1st ed.). Lippincott Williams & Wilkins. pp. 455. ISBN 978-0781765275.
  38. URL: http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain. Accessed on: 4 August 2010.
  39. URL: http://www.mayomedicallaboratories.com/test-catalog/Overview/80327. Accessed on: 8 August 2010.
  40. http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF
  41. Lefkowitch, Jay H. (2006). Anatomic Pathology Board Review (1st ed.). Saunders. pp. 681 (Q25). ISBN 978-1416025887.
  42. Humphrey, Peter A; Dehner, Louis P; Pfeifer, John D (2008). The Washington Manual of Surgical Pathology (1st ed.). Lippincott Williams & Wilkins. pp. 678. ISBN 978-0781765275.
  43. http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg
  44. URL: http://www.pathologyoutlines.com/topic/stainsalcianblue.html. Accessed on: 11 October 2012.
  45. URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF. Accessed on: 20 December 2011.
  46. 46.0 46.1 Pomerance, A.; Slavin, G.; McWatt, J. (Jan 1976). "Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology.". J Clin Pathol 29 (1): 22-6. PMID 55419.
  47. URL: http://www.polyrnd.com/products/reagent-assembly-kits/conventional/amyloid-stain---sulfated-alcian-blue-(sab).aspx. Accessed on: October 15, 2014.
  48. [1]
  49. 49.0 49.1 Modified Movat's Pentachrome Stain. University Penn Medicine. URL: http://www.med.upenn.edu/mcrc/histology_core/movat.shtml. Accessed on: January 29, 2009.
  50. Kunnath-Velayudhan S, Larsen BT, Coley SM, De Michele S, Santoriello D, Colby TV, Bhagat G, Saqi A (March 2021). "Masson Trichrome and Sulfated Alcian Blue Stains Distinguish Light Chain Deposition Disease From Amyloidosis in the Lung". Am J Surg Pathol 45 (3): 405–413. doi:10.1097/PAS.0000000000001593. PMID 33002919.
  51. ULR: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF. Accessed on: 2 November 2011.
  52. Perry JR, Bilbao JM, Gray T (1992). "Fatal basilar vasculopathy complicating bacterial meningitis". Stroke 23 (8): 1175–8. PMID 1636194. Free Full Text.
  53. Jones, DB.. "Nephrotic glomerulonephritis.". Am J Pathol 33 (2): 313-29. PMC 1934622. PMID 13402889. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1934622/.
  54. URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF. Accessed on: 19 May 2011.
  55. Rungby, J.; Kassem, M.; Eriksen, EF.; Danscher, G. (Jun 1993). "The von Kossa reaction for calcium deposits: silver lactate staining increases sensitivity and reduces background.". Histochem J 25 (6): 446-51. PMID 8360080.
  56. Fischer EG, Moore MJ, Lager DJ (October 2006). "Fabry disease: a morphologic study of 11 cases". Mod. Pathol. 19 (10): 1295–301. doi:10.1038/modpathol.3800634. PMID 16799480. http://www.nature.com/modpathol/journal/v19/n10/abs/3800634a.html.
  57. Nicholas, SB.; Basgen, JM.; Sinha, S. (2011). "Using stereologic techniques for podocyte counting in the mouse: shifting the paradigm.". Am J Nephrol 33 Suppl 1: 1-7. doi:10.1159/000327564. PMID 21659728.
  58. URL: http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html. Accessed on: 17 March 2011.
  59. URL: http://www.dermnetnz.org/doctors/dermatopathology/stains.html. Accessed on: 17 March 2011.
  60. Horobin RW, Walter KJ (1987). "Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears". Histochemistry 86 (3): 331–6. PMID 2437082. http://www.springerlink.com/content/r81x25451m841866/.
  61. URL: http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm. Accessed on: 4 January 2010.
  62. URL: http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm. Accessed on: 30 August 2012.
  63. Gaitanis, G.; Chasapi, V.; Velegraki, A. (Aug 2005). "Novel application of the masson-fontana stain for demonstrating Malassezia species melanin-like pigment production in vitro and in clinical specimens.". J Clin Microbiol 43 (8): 4147-51. doi:10.1128/JCM.43.8.4147-4151.2005. PMID 16081962.
  64. URL: http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html. Accessed on: 5 May 2010.
  65. URL: http://www.bris.ac.uk/vetpath/cpl/msb.html. Accessed on: 26 November 2010.
  66. URL: http://www.bris.ac.uk/vetpath/cpl/msb.html. Accessed on: 26 November 2010.
  67. "Picro-Mallory for Fibrin – Long Version". http://stainsfile.info/StainsFile/stain/fibrin/picro-mallory-1.htm. Retrieved 17 January 2011.
  68. URL: http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm. Accessed on: 3 January 2011.
  69. URL: http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvvg.htm. Accessed on: 3 January 2011.
  70. Miyamura H, Nakanuma Y, Kono N (December 1988). "Survey of copper granules in liver biopsy specimens from various liver abnormalities other than Wilson's disease and biliary diseases". Gastroenterol. Jpn. 23 (6): 633–8. PMID 2464523.
  71. URL: http://www.naika.or.jp/im2/42/10/14c.aspx. Accessed on: 24 January 2011.
  72. http://www.mayomedicallaboratories.com/test-catalog/Overview/9836. Accessed on: 24 January 2011.
  73. URL: http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat. Accessed on: 24 January 2011.
  74. Ghosh, AK.; Dasgupta, A.; Raha, K.; Jana, A.; Majumdar, DN. (Oct 1994). "Hepatic histology in chronic liver disease in hepatitis B surface antigen positive cases.". J Indian Med Assoc 92 (10): 333-5. PMID 7822848.
  75. URL: http://www.nottingham.ac.uk/pathology/protocols/shikata.html. Accessed on: 24 January 2011.
  76. GOMORI, G. - A rapid one-step trichrome stain. Am. J. Clin. Path. 20: 661-664, 1950.
  77. URL: https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm. Accessed on: 28 August 2015.

External links