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


=Where to start...=
=Where to start...=
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==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.
 
====Images====
<gallery>
Image:Cecal adenocarcinoma.jpg | Cecal adenocarcinoma. H&E stain. (WC)
Image:Basal cell carcinoma - high mag.jpg | [[Basal cell carcinoma]]. H&E stain. (WC)
</gallery>


==Haematoxylin phyloxin saffron stain==
==Haematoxylin phyloxin saffron stain==
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<gallery>  
<gallery>  
Image:Metanephric_adenoma_high_mag.jpg | Metanephric adenoma - PAS (WC)
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>
</gallery>


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*Light green = background.
*Light green = background.
*Washed-out light purple = Gram positive bacilli.
*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.
<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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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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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 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>
*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 stain.
*Auramine-rhodamine stain.


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Note:
Note:
*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>
*[[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.
**If the section is too thin... it doesn't work.


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Image:Cerebral_amyloid_angiopathy_-_very_high_mag.jpg | Congo red staining in [[cerebral amyloid angiopathy]]. (WC)
Image:Cerebral_amyloid_angiopathy_-_very_high_mag.jpg | Congo red staining in [[cerebral amyloid angiopathy]]. (WC)
</gallery>
</gallery>
==Thioflavin T stain==
==Thioflavin T stain==
===Use===
===Use===
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| Streptococcus pneumoniae, Staphylococcus aureus
| Streptococcus pneumoniae, Staphylococcus aureus
| Neisseria meningitidis, Moraxella catarrhalis  
| Neisseria meningitidis, Moraxella catarrhalis  
|
|}
|}


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<gallery>
<gallery>
Image:Globus_pallidus_and_putamen_-_very_low_mag.jpg | Globus pallidus and putamen - H&E-LFB. (WC)
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>
</gallery>
==Giemsa stain==
==Giemsa stain==
===Use===
===Use===
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===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==
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**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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Image:Liver_reticulin.jpg | [[Liver]]. Reticulin stain. (WC)
Image:Liver_reticulin.jpg | [[Liver]]. Reticulin stain. (WC)
Image:Hepatic_adenoma_high_mag_reticulin.jpg | [[Hepatic adenoma]]. 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>
</gallery>


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<gallery>
<gallery>
Image:Hemosiderosis_high_mag.jpg | Liver [[hemosiderosis]]. Prussian blue stain. (WC/Nephron)
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>
</gallery>


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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:
Notes:
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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===
*Stains 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.
*Uncommon.


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Image:Barrett's_mucosa,_higher_magnification,_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>
</gallery>
==Sodium sulphate-alcian blue stain==
:''Sulfated alcian blue'' (abbreviated '''SAB'') redirects here.
===Use===
*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===
*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)].


==Movat's stain==
==Movat's stain==
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====Images====
====Images====
<gallery>
<gallery>
Image:Cardiac_amyloidosis_very_high_mag_movat.jpg | Cardiac amyloidosis - Movat stain. (WC/Nephron)
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_-_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_-_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)
Image:Cystic medial degeneration - movat - high mag.jpg | Cystic medial degeneration - Movat stain - high mag. (WC/Nephron)
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*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>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>
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>
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==Hale's colloidal iron stain==
==Hale's colloidal iron stain==
===Use===
*[[Chromophobe renal cell carcinoma]] vs. [[renal oncocytoma]] - chromophobe [[renal cell carcinoma]] +ve.<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>


Notes:
{{Main|Hale's colloidal iron stain}}


===Interpretation===
==von Kossa stain==
*Blue (granular cytoplasmic) staining is positive.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</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>
Images:
*[http://www.ultrapath.org/oldsite/cases99/sep99/images/figure-3.jpg ChRCC Hale's colloidal iron (ultrapath.org)].<ref>URL: [http://www.ultrapath.org/oldsite/cases99/sep99/cotm9-2.html http://www.ultrapath.org/oldsite/cases99/sep99/cotm9-2.html]. Accessed on: 9 October 2011.</ref>
*[http://www.diagnosticpathology.org/content/4/1/21/figure/f1?highres=n ChRCC Hale's colloidal iron (diagnosticpathology.org)].
 
Notes:
*Often described as a "fastidious" (difficult/demanding) stain.<ref>URL: [http://www.merriam-webster.com/dictionary/fastidious?show=0&t=1319550566 http://www.merriam-webster.com/dictionary/fastidious?show=0&t=1319550566]. Accessed on: 25 October 2011.</ref>
**A few staff think this is a totally useless stain.<ref>ALS. On several occasions in 2009.</ref>


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


===Interpretation===
===Interpretation===
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*[[Cytopathology]].
*[[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>
*[[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===
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==Papanicolaou stain==
==Papanicolaou stain==
*Abbreviated ''Pap stain''.
*Abbreviated ''Pap stain''.
*Can be thought of as the [[H&E stain|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====
<gallery>
Image:Urine_citology_urothelial_carcinoma_2.jpg | Pap stain - [[urine cytology]] (WC)
</gallery>


==Fontana-Masson stain==  
==Fontana-Masson stain==  
*[[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''.
*[[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''.
*A type of silver stain.
{{Main|Fontana-Masson stain}}
 
Stains:
#Melanin.
#"Argentaffin granules" of the digestive tract.
#Pigment deposition due to minocycline treatment.<ref name=pmid15605966>{{Cite journal  | last1 = Patterson | first1 = JW. | last2 = Wilson | first2 = B. | last3 = Wick | first3 = MR. | last4 = Heath | first4 = C. | title = Hyperpigmented scar due to minocycline therapy. | journal = Cutis | volume = 74 | issue = 5 | pages = 293-8 | month = Nov | year = 2004 | doi =  | PMID = 15605966 }}</ref>
 
===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:
*[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm Fontana-Masson stain (vetmed.vt.edu)].


==Schmorl's stain==
==Schmorl's stain==
Line 697: 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>
Line 714: Line 728:


==Verhoeff-van Gieson stain==
==Verhoeff-van Gieson stain==
:''Verhoeff stain'' redirects here.
*[[AKA]] Elastic van Gieson stain, abbreviated ''EVG''.
*[[AKA]] Elastic van Gieson stain, abbreviated ''EVG''.
===General===
===General===
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*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==
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===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
*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.
*Red = nuclei.
*Light purple = background
*Bright red = mitochondria, red blood cells.
*??? = Copper associated protein.
 
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=

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.
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  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.
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