[Histonet] Brain Problems

[Tony Henwood]

The fixation time is too short.

Fix as long as you can (how about 2-3 days at least for brain?)

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Poole, Kimberly
Sent: Wednesday, 28 July 2010 12:28 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Brain Problems


Hi there,

 

I am currently using rat brain for histology but something is not working. I follow the protocol below for tissue processing.

 

Station

Solution

Routine (mins)

 

 

 

1

Formalin

10

2

Formalin

10

3

70% Ethanol

60

4

80% Ethanol

240

5

96% Ethanol

240

6

100% Ethanol

240

7

Xylene

120

8

Xylene

120

9

Xylene

120

10

Paraffin Wax

120

11

Paraffin Wax

180

12

Paraffin Wax

180

TOTAL TIME

 

27 hours 20 minutes

 

 

	

 

I embedded my brains and then cut them afterwards. When the brain section hits the water, it looks like a blob of fat on the water. Then it spreads reall, really slowly. If a cut is made to the paraffin the brain bursts open and it falls apart. Am I doing something wrong in the processing stage? Is the tissue dehydrated too much. Any feedback would be greatly appreciated.

Kimberly Poole B.Sc

Casualty Management Section | Section de la gestion des blessés

Defence Research and Development Canada Suffield | Recherche et développement pour la défense Canada Suffield 

Medicine Hat, AB, Canada T1A 8K6

kimberly.poole <@t> drdc-rddc.gc.ca <mailto:kimberly.poole <@t> drdc-rddc.gc.ca> 

Telephone | Téléphone 403-544-5347 / Facsimile | Télécopieur 403-544-4714 

Government of Canada | Gouvernement du Canada

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