Fixing cells - Revision history

Reshma P. Shetty at 18:42, 9 May 2007

Reshma P. Shetty — Wed, 09 May 2007 18:42:31 GMT

←Older revision		Revision as of 18:42, 9 May 2007
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	==External Links==		==External Links==
	[http://www.chemicon.com/techsupp/AcetoneMethod.asp Acetone Fixation protocol]		[http://www.chemicon.com/techsupp/AcetoneMethod.asp Acetone Fixation protocol]
		+
		+	[[Category:Protocol]] [[Category:Microscopy]]

Btarlow — Wed, 21 Mar 2007 05:57:09 GMT

'''Fixation'''

←Older revision		Revision as of 05:57, 21 March 2007
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	== '''Fixation''' ==		== '''Fixation''' ==

-	This can work well for fixing of mammalian cells in preparation for immunolabeling and microscopy. Depending on the epitope, different fixatives may be required; membrane-bound epitopes may do better without permeabilization.	+	This can work well for fixing of mammalian cells in preparation for immunolabeling and microscopy. Depending on the epitope, different fixatives may be required; membrane-bound epitopes may do better without permeabilization. On the other hand, some antibodies are only able to recognize a protein epitope in its native form and will only work on frozen tissues because formaldehyde cross-linking denatures the protein.

	'''Reagents'''		'''Reagents'''
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	For further information about fixing cells, see [[Immunocytochemistry]].		For further information about fixing cells, see [[Immunocytochemistry]].
		+
		+	==External Links==
		+	[http://www.chemicon.com/techsupp/AcetoneMethod.asp Acetone Fixation protocol]

Etchevers — Mon, 04 Dec 2006 12:13:51 GMT

←Older revision		Revision as of 12:13, 4 December 2006
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-	== '''~~Formaldehyde~~ Fixation''' ==	+	== '''Fixation''' ==

-	*This ~~works~~ well for ~~fixation~~ of mammalian cells in preparation for immunolabeling and microscopy	+	This can work well for fixing of mammalian cells in preparation for immunolabeling and microscopy. Depending on the epitope, different fixatives may be required; membrane-bound epitopes may do better without permeabilization.

	'''Reagents'''		'''Reagents'''

-	-Fixation Solution: 4% Formaldehyde in PBS	+	-Fixation Solution: 4% Formaldehyde in phosphate buffered saline (PBS) solution, pH 7.4 - 7.6 (verify)

	-Permeabilization Solution: 0.1% Triton X-100 in PBS		-Permeabilization Solution: 0.1% Triton X-100 in PBS
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	2) For permeabilization, remove Formaldehyde Solution, and incubate cells in Permeabilization Solution for 5 minutes at room temperature.		2) For permeabilization, remove Formaldehyde Solution, and incubate cells in Permeabilization Solution for 5 minutes at room temperature.
		+
		+	3) Rinse in PBS before proceeding.


	For further information about fixing cells, see [[Immunocytochemistry]].		For further information about fixing cells, see [[Immunocytochemistry]].

Holmesbr — Thu, 15 Jun 2006 17:21:44 GMT

←Older revision		Revision as of 17:21, 15 June 2006
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	2) For permeabilization, remove Formaldehyde Solution, and incubate cells in Permeabilization Solution for 5 minutes at room temperature.		2) For permeabilization, remove Formaldehyde Solution, and incubate cells in Permeabilization Solution for 5 minutes at room temperature.
		+
		+
		+	For further information about fixing cells, see [[Immunocytochemistry]].

Holmesbr — Wed, 14 Jun 2006 19:17:54 GMT

←Older revision		Revision as of 19:17, 14 June 2006
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	== '''Formaldehyde Fixation''' ==		== '''Formaldehyde Fixation''' ==

-	***This works well for fixation of mammalian cells in preparation for immunolabeling and microscopy	+	*This works well for fixation of mammalian cells in preparation for immunolabeling and microscopy

	'''Reagents'''		'''Reagents'''

Bbelmont — Sun, 26 Feb 2006 06:13:35 GMT

←Older revision		Revision as of 06:13, 26 February 2006
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	== '''Formaldehyde Fixation''' ==		== '''Formaldehyde Fixation''' ==

-	***This works well for fixation of mammalian cells in preparation for ~~immunolabeleing~~ and microscopy	+	***This works well for fixation of mammalian cells in preparation for immunolabeling and microscopy

	'''Reagents'''		'''Reagents'''

Bbelmont — Sun, 26 Feb 2006 06:13:12 GMT

←Older revision		Revision as of 06:13, 26 February 2006
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	'''Reagents'''		'''Reagents'''
		+
	-Fixation Solution: 4% Formaldehyde in PBS		-Fixation Solution: 4% Formaldehyde in PBS

Bbelmont — Sun, 26 Feb 2006 06:13:01 GMT

←Older revision		Revision as of 06:13, 26 February 2006
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	'''Reagents'''		'''Reagents'''
	-Fixation Solution: 4% Formaldehyde in PBS		-Fixation Solution: 4% Formaldehyde in PBS
		+
	-Permeabilization Solution: 0.1% Triton X-100 in PBS		-Permeabilization Solution: 0.1% Triton X-100 in PBS

Bbelmont — Sun, 26 Feb 2006 06:12:49 GMT

←Older revision		Revision as of 06:12, 26 February 2006
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-
	== '''Formaldehyde Fixation''' ==		== '''Formaldehyde Fixation''' ==

	***This works well for fixation of mammalian cells in preparation for immunolabeleing and microscopy		***This works well for fixation of mammalian cells in preparation for immunolabeleing and microscopy

-	~~1) Incubate cells in ~~~4% Formaldehyde ~~solution (~~in PBS) for 10-15 minutes at room temperature.	+	'''Reagents'''
		+	-Fixation Solution: 4% Formaldehyde in PBS
		+	-Permeabilization Solution: 0.1% Triton X-100 in PBS
		+
		+	'''Protocol'''
		+
		+	1) For fixation, incubate cells in Formaldehyde Solution for 10-15 minutes at room temperature.

-	2) For permeabilization ~~of cells~~, remove ~~formaldehyde solution~~, and incubate cells in ~~0.1% Triton X-100 (in PBS)~~ for 5 minutes at room temperature~~. There is no need to wash between fixation and permeabilization steps~~.	+	2) For permeabilization, remove Formaldehyde Solution, and incubate cells in Permeabilization Solution for 5 minutes at room temperature.

Bbelmont — Sun, 26 Feb 2006 06:11:02 GMT

←Older revision		Revision as of 06:11, 26 February 2006
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-	'''Formaldehyde Fixation'''	+
		+	== '''Formaldehyde Fixation''' ==
		+
	***This works well for fixation of mammalian cells in preparation for immunolabeleing and microscopy		***This works well for fixation of mammalian cells in preparation for immunolabeleing and microscopy