FW: [Histonet] Leder Stain

[Edmondson David (RBV) NHS Christie Tr]

Hi
Have no idea whose name should be attributed to this but..
Slides to water.
We have 30mg of Fast Blue BB base and 6mg of alpha Naphthyl Chloroacetate The first dissolved in 0.2M pH 7.6 TrisMalate buffer, the second in a few drops of Dimethyl Formamide and then added to the buffer. All is mixed and the slides immersed for 20-30mins.  37deg C is fine but go for a shorter time to avoid background. 
Counterstain say Neutral Red, rinse then dehydrate rapidly and mount.  Take too long in alcohol or waiting to be mounted may abolish the staining.
Check hazards.  Do risk assessment 

To save calculations, could look in Bancroft and Stevens appendix for buffer table.where dissolve 2.42g TRIS + 2.32g Maleic acid in 100ml of water
then 0.8g NaOH in 100ml of water
25ml of first plus 29ml of alkali give pH7.6

Dave
Christie Hosp
Manchester UK
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Louro,
Pedro
Sent: 30 March 2005 15:00
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Leder Stain


I'm looking for information on "Leder Stain" (chloracetate esterase
activity)...looking to stain neutrophils on FFPE mouse tissue
If someone can supply me with the recipe or the ingredients would be great

Thanks 

Pedro


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