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Immunohistochemical staining (view source)
Revision as of 11:49, 30 November 2020
, 11:49, 30 November 2020→Signal assessment: Update
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**Less prone to false positives. | **Less prone to false positives. | ||
***Negative controls not needed or infrequently required.<ref name=pmid24714041>{{Cite journal | last1 = Torlakovic | first1 = EE. | last2 = Francis | first2 = G. | last3 = Garratt | first3 = J. | last4 = Gilks | first4 = B. | last5 = Hyjek | first5 = E. | last6 = Ibrahim | first6 = M. | last7 = Miller | first7 = R. | last8 = Nielsen | first8 = S. | last9 = Petcu | first9 = EB. | title = Standardization of negative controls in diagnostic immunohistochemistry: recommendations from the international ad hoc expert panel. | journal = Appl Immunohistochem Mol Morphol | volume = 22 | issue = 4 | pages = 241-52 | month = Apr | year = 2014 | doi = 10.1097/PAI.0000000000000069 | PMID = 24714041 }}</ref> | ***Negative controls not needed or infrequently required.<ref name=pmid24714041>{{Cite journal | last1 = Torlakovic | first1 = EE. | last2 = Francis | first2 = G. | last3 = Garratt | first3 = J. | last4 = Gilks | first4 = B. | last5 = Hyjek | first5 = E. | last6 = Ibrahim | first6 = M. | last7 = Miller | first7 = R. | last8 = Nielsen | first8 = S. | last9 = Petcu | first9 = EB. | title = Standardization of negative controls in diagnostic immunohistochemistry: recommendations from the international ad hoc expert panel. | journal = Appl Immunohistochem Mol Morphol | volume = 22 | issue = 4 | pages = 241-52 | month = Apr | year = 2014 | doi = 10.1097/PAI.0000000000000069 | PMID = 24714041 }}</ref> | ||
===Signal assessment=== | |||
*Manual (Morphology) vs. automated (Speed) counting. | |||
*Choice of proper chromogen. | |||
**Dynamic range of DAB is 1-2logs vs. fluorescent probes 2-3logs. | |||
**Protein expression range can be up to 4 logs in gene amplification (information is missed in IHC). | |||
**FastRed stains are more suited in melanocytic tumors than DAB. | |||
*Use of established cutoffs for "intensity" and for "positive" staining. | |||
*Visual pattern recogniton vs. detecting spatial subtle changes. | |||
*Detecting differences at low intensity with human eye is less accurate. | |||
==Quality control== | ==Quality control== | ||
This is an evolving area in pathology that has been ignored for a surprisingly long time. | This is an evolving area in pathology that has been ignored for a surprisingly long time. | ||
It is touched upon the in the ''[[quality]]'' article in the ''[[Quality#Immunohistochemistry|immunohistochemistry]]'' section. | It is touched upon the in the ''[[quality]]'' article in the ''[[Quality#Immunohistochemistry|immunohistochemistry]]'' section. | ||
There are at least 62 pre-analytical variables to be considered, that may affect staining results.<ref>{{Cite journal | last1 = Engel | first1 = KB. | last2 = Moore | first2 = HM. | title = Effects of preanalytical variables on the detection of proteins by immunohistochemistry in formalin-fixed, paraffin-embedded tissue. | journal = Arch Pathol Lab Med | volume = 135 | issue = 5 | pages = 537-43 | month = May | year = 2011 | doi = 10.1043/2010-0702-RAIR.1 | PMID = 21526952 }}</ref> | |||
==Interpretation== | ==Interpretation== | ||
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*Glypican-3. | *Glypican-3. | ||
**[[Hepatocellular carcinoma]] (HCC) stains with glypican 3, while [[liver]] with dysplastic changes and/or [[cirrhosis|cirrhotic changes]] does not.<ref>{{cite journal | author=Anatelli F, Chuang ST, Yang XJ, Wang HL. |title=Value of glypican 3 immunostaining in the diagnosis of hepatocellular carcinoma on needle biopsy |journal=Am J Clin Pathol. |volume=130 |issue= 2 |pages= 219-23? |year= 2008 |pmid= 18628090 |doi= 10.1309/WMB5PX57Y4P8QCTY }}</ref> | **[[Hepatocellular carcinoma]] (HCC) stains with glypican 3, while [[liver]] with dysplastic changes and/or [[cirrhosis|cirrhotic changes]] does not.<ref>{{cite journal | author=Anatelli F, Chuang ST, Yang XJ, Wang HL. |title=Value of glypican 3 immunostaining in the diagnosis of hepatocellular carcinoma on needle biopsy |journal=Am J Clin Pathol. |volume=130 |issue= 2 |pages= 219-23? |year= 2008 |pmid= 18628090 |doi= 10.1309/WMB5PX57Y4P8QCTY }}</ref> | ||
*HepPar-1 (hepatocytes paraffin antibody 1) - labels hepatocellular mitochondria.<ref name=pmid12502967>The diagnostic value of hepatocyte paraffin antibody 1 in differentiating hepatocellular neoplasms from nonhepatic tumors: a review. Lamps LW, Folpe AL. Adv Anat Pathol. 2003 Jan;10(1):39-43. Review. PMID 12502967.</ref> | *[[HepPar-1]] (hepatocytes paraffin antibody 1) - labels hepatocellular mitochondria.<ref name=pmid12502967>The diagnostic value of hepatocyte paraffin antibody 1 in differentiating hepatocellular neoplasms from nonhepatic tumors: a review. Lamps LW, Folpe AL. Adv Anat Pathol. 2003 Jan;10(1):39-43. Review. PMID 12502967.</ref> | ||
[[HCC]] vs. [[cholangiocarcinoma]]: | [[HCC]] vs. [[cholangiocarcinoma]]: | ||