Tissue fixation - Revision history

Mark: my contribution from ganfyd.org (now inactive), slightly modified

2025-10-12T19:48:51Z

my contribution from ganfyd.org (now inactive), slightly modified

← Older revision		Revision as of 19:48, 12 October 2025
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	*May be written (incorrectly) as "formulin".		*May be written (incorrectly) as "formulin".
	*Formaldehyde + methanol.		*Formaldehyde + methanol.

			Formaldehyde is often supplied as a 37% w/v solution. To prevent polymerisation of formaldehyde, methanol is added. The resulting solution is often referred to as formalin. This can be further diluted and buffered to produce 10% formalin (equivalent to 4% formaldehyde). This concentration is often used as a fixative and preservative for histological specimens. It gives tissues a firmer texture, making subsequent cut-up easier and also stops autolysis and necrosis of the tissue by inactivating micro-organisms and proteolytic enzymes. Poorly fixed tissue stains unreliably and certain features may be lost. Mitotic figures, for instance, are easily lost in poorly fixed tissue. Formalin is also used to preserve prosected and dissected cadavers. As a rule of thumb, fixation occurs at about 1 mm per hour, but the reality is more complex as fixation lags behind diffusion and fixation also changes the tissue properties, slowing diffusion.<ref name="pmid22483550">{{cite journal \|vauthors=Buesa RJ, Peshkov MV \|title=How much formalin is enough to fix tissues? \|journal=Ann Diagn Pathol \|volume=16 \|issue=3 \|pages=202–9 \|date=June 2012 \|pmid=22483550 \|doi=10.1016/j.anndiagpath.2011.12.003 \|url=}}</ref>

			Formalin cross-links sections of the peptide chains of proteins as well as nucleic acids. The effect on the former alters epitopes, which means that antibodies designed for fresh tissue may not work with fixed tissue. The cross-linking process can be partially reversed with various methods of antigen retrieval. The effect on nucleic acids is to reduce DNA quality.

	=Fixing marking dye=		=Fixing marking dye=

Michael: /* See also */

2017-10-19T13:23:04Z

Michael: /* See also */

2014-09-26T17:37:45Z

Michael: /* Tissue fixation */

2014-09-26T17:36:01Z

Tissue fixation

← Older revision		Revision as of 17:36, 26 September 2014
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	\|-		\|-
	\| B5		\| B5
	\| used for lymphoma, superior morphology - but toxic, <br>not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>		\| used for [[lymphoma]], superior morphology - but toxic, <br>not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>
	\|-		\|-
	\| IBF fixative		\| IBF fixative
	\| used ~~for prostate biopsies~~ - gives better nuclear detail<ref name=pmid16390230>{{Cite journal \| last1 = Trpkov \| first1 = K. \| last2 = Renault \| first2 = P. \| last3 = Yilmaz \| first3 = A. \| last4 = Ali-Ridha \| first4 = N. \| title = IBF as a formalin substitute fixative in prostate biopsy pathology. \| journal = Arch Pathol Lab Med \| volume = 130 \| issue = 1 \| pages = 13-4 \| month = Jan \| year = 2006 \| doi = 10.1043/1543-2165(2006)130[13b:IAAFSF]2.0.CO;2 \| PMID = 16390230 \|URL = http://www.archivesofpathology.org/doi/full/10.1043/1543-2165(2006)130%5B13b%3AIAAFSF%5D2.0.CO%3B2 }}</ref>		\| used bone marrow and lymphatic tissue;<ref>URL: [http://www.leicabiosystems.com/specimen-preparation/consumables/reagents-solutions/fixatives/details/product/ibf-tissue-fixative-1/ http://www.leicabiosystems.com/specimen-preparation/consumables/reagents-solutions/fixatives/details/product/ibf-tissue-fixative-1/]. Accessed on: 26 September 2014.</ref> gives better nuclear detail in prostate biopsies compared to formalin<ref name=pmid16390230>{{Cite journal \| last1 = Trpkov \| first1 = K. \| last2 = Renault \| first2 = P. \| last3 = Yilmaz \| first3 = A. \| last4 = Ali-Ridha \| first4 = N. \| title = IBF as a formalin substitute fixative in prostate biopsy pathology. \| journal = Arch Pathol Lab Med \| volume = 130 \| issue = 1 \| pages = 13-4 \| month = Jan \| year = 2006 \| doi = 10.1043/1543-2165(2006)130[13b:IAAFSF]2.0.CO;2 \| PMID = 16390230 \|URL = http://www.archivesofpathology.org/doi/full/10.1043/1543-2165(2006)130%5B13b%3AIAAFSF%5D2.0.CO%3B2 }}</ref>
	\|}		\|}

Michael: /* Tissue fixation */

2014-09-26T17:32:50Z

Tissue fixation

← Older revision		Revision as of 17:32, 26 September 2014
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	\|-		\|-
	\| IBF fixative		\| IBF fixative
	\| used for prostate biopsies - gives better nuclear detail<ref name=pmid16390230>{{Cite journal \| last1 = Trpkov \| first1 = K. \| last2 = Renault \| first2 = P. \| last3 = Yilmaz \| first3 = A. \| last4 = Ali-Ridha \| first4 = N. \| title = IBF as a formalin substitute fixative in prostate biopsy pathology. \| journal = Arch Pathol Lab Med \| volume = 130 \| issue = 1 \| pages = 13-4 \| month = Jan \| year = 2006 \| doi = 10.1043/1543-2165(2006)130[13b:IAAFSF]2.0.CO;2 \| PMID = 16390230 }}</ref>		\| used for prostate biopsies - gives better nuclear detail<ref name=pmid16390230>{{Cite journal \| last1 = Trpkov \| first1 = K. \| last2 = Renault \| first2 = P. \| last3 = Yilmaz \| first3 = A. \| last4 = Ali-Ridha \| first4 = N. \| title = IBF as a formalin substitute fixative in prostate biopsy pathology. \| journal = Arch Pathol Lab Med \| volume = 130 \| issue = 1 \| pages = 13-4 \| month = Jan \| year = 2006 \| doi = 10.1043/1543-2165(2006)130[13b:IAAFSF]2.0.CO;2 \| PMID = 16390230 \|URL = http://www.archivesofpathology.org/doi/full/10.1043/1543-2165(2006)130%5B13b%3AIAAFSF%5D2.0.CO%3B2 }}</ref>


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Michael: /* Tissue fixation */

2014-09-26T17:30:14Z

Tissue fixation

← Older revision		Revision as of 17:30, 26 September 2014
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	\| used for lymphoma, superior morphology - but toxic, <br>not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>		\| used for lymphoma, superior morphology - but toxic, <br>not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>
	\|-		\|-
			\| IBF fixative
			\| used for prostate biopsies - gives better nuclear detail<ref name=pmid16390230>{{Cite journal \| last1 = Trpkov \| first1 = K. \| last2 = Renault \| first2 = P. \| last3 = Yilmaz \| first3 = A. \| last4 = Ali-Ridha \| first4 = N. \| title = IBF as a formalin substitute fixative in prostate biopsy pathology. \| journal = Arch Pathol Lab Med \| volume = 130 \| issue = 1 \| pages = 13-4 \| month = Jan \| year = 2006 \| doi = 10.1043/1543-2165(2006)130[13b:IAAFSF]2.0.CO;2 \| PMID = 16390230 }}</ref>


	\|}		\|}

Michael: /* Tissue fixation */

2014-02-04T15:29:33Z

Tissue fixation

← Older revision		Revision as of 15:29, 4 February 2014
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	\| ???		\| ???
	\|-		\|-
	\| Glutaraldehyde		\| Glutaraldehyde<ref name=pmid8787969>{{Cite journal \| last1 = Prentø \| first1 = P. \| title = Glutaraldehyde for electron microscopy: a practical investigation of commercial glutaraldehydes and glutaraldehyde-storage conditions. \| journal = Histochem J \| volume = 27 \| issue = 11 \| pages = 906-13 \| month = Nov \| year = 1995 \| doi = \| PMID = 8787969 }}</ref>
	\| [[electron microscopy]]		\| [[electron microscopy]]
	\|-		\|-

Michael: /* Tissue fixation */

2012-03-28T14:03:10Z

Tissue fixation

← Older revision		Revision as of 14:03, 28 March 2012
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	\|-		\|-
	\| B5		\| B5
	\| used for lymphoma, superior morphology - but toxic, not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>		\| used for lymphoma, superior morphology - but toxic, <br>not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>
	\|-		\|-
	\|}		\|}

Michael: /* Tissue fixation */ more

2012-03-28T13:42:00Z

Tissue fixation: more

← Older revision		Revision as of 13:42, 28 March 2012
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	\|-		\|-
	\| Glutaraldehyde		\| Glutaraldehyde
	\| [[electron microscopy]] ~~???~~		\| [[electron microscopy]]
	\|-		\|-
	\| Ethanol		\| Ethanol
	\| [[cytopathology]] ~~???~~		\| [[cytopathology]]
	\|-		\|-
	\| Carnoy		\| Carnoy
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	\| Bouin		\| Bouin
	\| toxic ???		\| toxic ???
			\|-
			\| B5
			\| used for lymphoma, superior morphology - but toxic, not good for IHC & DNA analysis<ref>{{Cite journal \| last1 = Bonds \| first1 = LA. \| last2 = Barnes \| first2 = P. \| last3 = Foucar \| first3 = K. \| last4 = Sever \| first4 = CE. \| title = Acetic acid-zinc-formalin: a safe alternative to B-5 fixative. \| journal = Am J Clin Pathol \| volume = 124 \| issue = 2 \| pages = 205-11 \| month = Aug \| year = 2005 \| doi = 10.1309/29DA-CY9K-BHNW-4BG6 \| PMID = 16040290 }}</ref>
			\|-
	\|}		\|}

Michael: more

2012-03-27T18:29:14Z

← Older revision		Revision as of 18:29, 27 March 2012
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	Pathologist have a great lifestyle 'cause tissue takes long to fix; the penetration of tissue by formalin is 1 mm/hour.<ref>Gross rounds. 14 August 2009.</ref>		Pathologist have a great lifestyle 'cause tissue takes long to fix; the penetration of tissue by formalin is 1 mm/hour.<ref>Gross rounds. 14 August 2009.</ref>

	==~~Fixing marking dye~~==		==Ratio==
	~~To fix marking dye:~~		The dictum is:<ref>URL: [http://www.cap.org/apps/docs/committees/cancer/cancer_protocols/2011/UvealMelanom_11protocol.pdf http://www.cap.org/apps/docs/committees/cancer/cancer_protocols/2011/UvealMelanom_11protocol.pdf]. Accessed on: 27 March 2012.</ref>
	*Formal-acetic alcohol (FAA):<ref>URL: [http://www.~~brunelmicroscopes~~.~~co.uk~~/~~acatalog~~/~~Solvents___Reagents~~.~~html~~ http://www.~~brunelmicroscopes~~.~~co.uk~~/~~acatalog~~/~~Solvents___Reagents~~.~~html~~]. Accessed on: ~~5 January 2011~~.</ref>		*''The volume of fixative should be 10x the volume of specimen.''
	*~~Bouin~~'~~s solution~~.

	==Tissue fixation==		==Tissue fixation==
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	*Formaldehyde + methanol.		*Formaldehyde + methanol.

	=~~Dye fixation~~=		=Fixing marking dye=
			To fix marking dye:
			*Formal-acetic alcohol (FAA):<ref>URL: [http://www.brunelmicroscopes.co.uk/acatalog/Solvents___Reagents.html http://www.brunelmicroscopes.co.uk/acatalog/Solvents___Reagents.html]. Accessed on: 5 January 2011.</ref>
			*Bouin's solution.

	==Formal-acetic-alcohol==		==Formal-acetic-alcohol==
	General:		General:

← Older revision		Revision as of 13:23, 19 October 2017
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	*[[Basics]].		*[[Basics]].
	*[[Histology artifacts]].		*[[Histology artifacts]].
			*[[Formalin-fixed paraffin-embedded]].

	=References=		=References=

← Older revision		Revision as of 17:37, 26 September 2014
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	=See also=		=See also=
	*[[Basics]].		*[[Basics]].
			*[[Histology artifacts]].

	=References=		=References=