Immunohistochemical staining - Revision history

Jensflorian: /* Signal assessment */ Update

2020-11-30T11:49:57Z

Signal assessment: Update

← Older revision		Revision as of 11:49, 30 November 2020
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	===Signal assessment===		===Signal assessment===
	*Manual vs. automated counting.		*Manual (Morphology) vs. automated (Speed) counting.
	*Choice of proper chromogen.		*Choice of proper chromogen.
	**Dynamic range of DAB is 1-2logs vs. fluorescent probes 2-3logs.		**Dynamic range of DAB is 1-2logs vs. fluorescent probes 2-3logs.

Jensflorian: /* Signal assessment */ Update

2020-11-30T11:48:27Z

Signal assessment: Update

← Older revision		Revision as of 11:48, 30 November 2020
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	===Signal assessment===		===Signal assessment===
	*Manual ~~vas~~ automated.		*Manual vs. automated counting.
	*Choice of chromogen		*Choice of proper chromogen.
	**Dynamic range of DAB is 1-2logs vs. fluorescent probes 2-3logs.		**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).		**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.		**FastRed stains are more suited in melanocytic tumors than DAB.
	*~~Cutoffs~~ for "intensity" and for "positive" staining.		*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==

Jensflorian: /* Theory */ Update

2020-11-30T11:42:54Z

Theory: Update

← Older revision		Revision as of 11:42, 30 November 2020
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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 vas automated.
			*Choice of 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.
			*Cutoffs for "intensity" and for "positive" staining.

	==Quality control==		==Quality control==

Michael: wikify

2019-11-22T16:23:29Z

wikify

← Older revision		Revision as of 16:23, 22 November 2019
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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]]:

Jensflorian: /* Quality control */ pre-analytical variables

2018-07-02T12:31:49Z

Quality control: pre-analytical variables

← Older revision		Revision as of 12:31, 2 July 2018
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	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==

Jensflorian: /* Utility */ +coons

2018-07-02T12:19:29Z

Utility: +coons

← Older revision		Revision as of 12:19, 2 July 2018
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	#Mutation specific antibodies, eg. [[IDH-1]] R132H.		#Mutation specific antibodies, eg. [[IDH-1]] R132H.

	Method was introduced in 1941 by Coons.<{{Cite journal \| last1 = Coons \| first1 = AH. \| title = The development of immunohistochemistry. \| journal = Ann N Y Acad Sci \| volume = 177 \| issue = \| pages = 5-9 \| month = Jun \| year = 1971 \| doi = \| PMID = 4400556 }}</ref>		Method was introduced in 1941 by Coons.<ref>{{Cite journal \| last1 = Coons \| first1 = AH. \| title = The development of immunohistochemistry. \| journal = Ann N Y Acad Sci \| volume = 177 \| issue = \| pages = 5-9 \| month = Jun \| year = 1971 \| doi = \| PMID = 4400556 }}</ref>

	==Theory==		==Theory==

Jensflorian: /* Utility */ +Coons

2018-07-02T12:19:11Z

Utility: +Coons

← Older revision		Revision as of 12:19, 2 July 2018
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	#Prognostic markers, e.g. ERBB2 (HER2).		#Prognostic markers, e.g. ERBB2 (HER2).
	#Proving clonality - in the context of hematologic malignancies.		#Proving clonality - in the context of hematologic malignancies.
			#Mutation specific antibodies, eg. [[IDH-1]] R132H.

			Method was introduced in 1941 by Coons.<{{Cite journal \| last1 = Coons \| first1 = AH. \| title = The development of immunohistochemistry. \| journal = Ann N Y Acad Sci \| volume = 177 \| issue = \| pages = 5-9 \| month = Jun \| year = 1971 \| doi = \| PMID = 4400556 }}</ref>

	==Theory==		==Theory==

Jensflorian: /* Breast markers */ eikify ER

2018-01-25T09:36:42Z

Breast markers: eikify ER

← Older revision		Revision as of 09:36, 25 January 2018
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	===Breast markers===		===Breast markers===
	*[[GCDFP-15]] ([[AKA]] BRST-2) -- specific, but NOT sensitive.		*[[GCDFP-15]] ([[AKA]] BRST-2) -- specific, but NOT sensitive.
	*ER (estrogen receptor) - in normal [[breast]].		*[[Estrogen receptor\|ER]] (estrogen receptor) - in normal [[breast]].
	*PR (progesterone receptor) - in normal breast.		*PR (progesterone receptor) - in normal breast.
	*HER2/neu - pathological, assoc. with worse prognosis.		*HER2/neu - pathological, assoc. with worse prognosis.

Michael: /* Interpretation */ tweak

2018-01-08T19:07:45Z

Interpretation: tweak

← Older revision		Revision as of 19:07, 8 January 2018
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	In cases where the morphology is unclear, it is judicious to have two or more immunostains that support the diagnosis, and negative stains for important entities in the differential diagnosis.		In cases where the morphology is unclear, it is judicious to have two or more immunostains that support the diagnosis, and negative stains for important entities in the differential diagnosis.

	Publications with contradicting ~~result~~ are not uncommon. Differences arise from the ~~interpretation~~, processing protocol ~~and~~ antibody clone.		Publications with contradicting results are not uncommon. Differences can arise from the fixation, processing protocol, antibody clone and interpretation.

	According to Galloway, one third pathologists substantially overestimate the diagnostic significance of unexpected immunohistochemical staining results.<ref name=pmid21660231>{{Cite journal \| last1 = Galloway \| first1 = M. \| title = Base-rate error in the interpretation of immunohistochemistry. \| journal = Patholog Res Int \| volume = 2011 \| issue = \| pages = 636495 \| month = \| year = 2011 \| doi = 10.4061/2011/636495 \| PMID = 21660231 }}</ref>		According to Galloway, one third pathologists substantially overestimate the diagnostic significance of unexpected immunohistochemical staining results.<ref name=pmid21660231>{{Cite journal \| last1 = Galloway \| first1 = M. \| title = Base-rate error in the interpretation of immunohistochemistry. \| journal = Patholog Res Int \| volume = 2011 \| issue = \| pages = 636495 \| month = \| year = 2011 \| doi = 10.4061/2011/636495 \| PMID = 21660231 }}</ref>

Michael: /* Signaling systems */

2017-07-07T13:50:29Z

Signaling systems

← Older revision		Revision as of 13:50, 7 July 2017
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	**Older.<ref>{{Cite journal \| last1 = Vosse \| first1 = BA. \| last2 = Seelentag \| first2 = W. \| last3 = Bachmann \| first3 = A. \| last4 = Bosman \| first4 = FT. \| last5 = Yan \| first5 = P. \| title = Background staining of visualization systems in immunohistochemistry: comparison of the Avidin-Biotin Complex system and the EnVision+ system. \| journal = Appl Immunohistochem Mol Morphol \| volume = 15 \| issue = 1 \| pages = 103-7 \| month = Mar \| year = 2007 \| doi = \| PMID = 17536316 }}</ref>		**Older.<ref>{{Cite journal \| last1 = Vosse \| first1 = BA. \| last2 = Seelentag \| first2 = W. \| last3 = Bachmann \| first3 = A. \| last4 = Bosman \| first4 = FT. \| last5 = Yan \| first5 = P. \| title = Background staining of visualization systems in immunohistochemistry: comparison of the Avidin-Biotin Complex system and the EnVision+ system. \| journal = Appl Immunohistochem Mol Morphol \| volume = 15 \| issue = 1 \| pages = 103-7 \| month = Mar \| year = 2007 \| doi = \| PMID = 17536316 }}</ref>
	**May suffer from endogenous avidin-biotin activity.<ref name=pmid17536316>{{Cite journal \| last1 = Vosse \| first1 = BA. \| last2 = Seelentag \| first2 = W. \| last3 = Bachmann \| first3 = A. \| last4 = Bosman \| first4 = FT. \| last5 = Yan \| first5 = P. \| title = Background staining of visualization systems in immunohistochemistry: comparison of the Avidin-Biotin Complex system and the EnVision+ system. \| journal = Appl Immunohistochem Mol Morphol \| volume = 15 \| issue = 1 \| pages = 103-7 \| month = Mar \| year = 2007 \| doi = \| PMID = 17536316 }}</ref>		**May suffer from endogenous avidin-biotin activity.<ref name=pmid17536316>{{Cite journal \| last1 = Vosse \| first1 = BA. \| last2 = Seelentag \| first2 = W. \| last3 = Bachmann \| first3 = A. \| last4 = Bosman \| first4 = FT. \| last5 = Yan \| first5 = P. \| title = Background staining of visualization systems in immunohistochemistry: comparison of the Avidin-Biotin Complex system and the EnVision+ system. \| journal = Appl Immunohistochem Mol Morphol \| volume = 15 \| issue = 1 \| pages = 103-7 \| month = Mar \| year = 2007 \| doi = \| PMID = 17536316 }}</ref>
			***Higher false positive rates than with polymer based methods.
	*Polymer based methods.		*Polymer based methods.
	**Newer.		**Newer.