[Histonet] blotches in sections[Scanned]

[Kemlo Rogerson]

Ooops sorry, didn't read Tony's reply. I concur.

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194
 

-----Original Message-----
From: Tony Henwood [mailto:AnthonyH <@t> chw.edu.au] 
Sent: 25 November 2004 21:50
To: 'Pool, Madeline'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] blotches in sections[Scanned]

Madeline,

My first impression is that there is water on the sections. You may possibly
have to prolong your dehydration and clearing prior to coverslipping. You
could decoverslip, remove the mountant in xylene and complete the
dehydration in ethanol and then (via xylene or its substitute) recoverslip.
Regards

Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318





-----Original Message-----
From: Pool, Madeline [mailto:mpool <@t> ohri.ca] 
Sent: Friday, 26 November 2004 6:15 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] blotches in sections


I'm cutting 10 um sections from paraffin embedded mouse brain, spinal
cord, and optic nerve and staining them with Luxol Fast Blue. In the
brain sections, I'm seeing some splotchy darker background. I'm
wondering if maybe I didn't dry the slides enough after sectioning or
maybe heated it too much or something else entirely. If anyone has seen
things like this before and can tell me what I should do to fix it,
please let me know. I've put a picture of a section up at:
http://members.rogers.com/madeline.pool/brain.jpg (Don't worry about the
tearing, I was testing out bad sections.)

Thanks in advance,
Madeline

Madeline Pool
Ottawa Health Research Institute
501 Smyth Road
Ottawa, Ontario
K1H 8L6
(613)739-6634
fax: (613)737-8023

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