[Histonet] Embryo sticking in InsituPro Run

[Grant, Debra]

Hi Jimmy,

We use 1% NaOH (Sodium Hydroxide) 6 hours to overnight, then we rinse 3x with distilled water, then soak them in distilled H20 overnight, then rinse 2x with distilled H20, then dry them in an oven @ 60 degrees Celsius, and rinse in 100% EtOH then dry again in the oven @ 60 degrees Celsius. Usually we can use the columns until the filters pop out from the columns, approximately 3-4 times. Also, if you use tween 20 at 0.1% in all hybridization solutions you shouldn't have this problem with the embryos sticking to the columns.

Debby Grant
Research Technician II
Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th Street
Kansas City, MO 64110
drg <@t> stowers-institute.org
 


-----Original Message-----
From: Jimmy Lao [mailto:lao_ji <@t> yahoo.com]
Sent: Tuesday, October 28, 2003 4:27 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Embryo sticking in InsituPro Run


Hi
Any one can help preventing embryo sticking to the
Column during InsituPro run? InsituPro is automated
machine for whole mount embryo in situ hybridization. 
It did not happen when I used fresh column. But it did
in the 2nd time using column.  I guess there are
something staying in the column after first use, even
I treated them with H2O2 overnight and 4%PFA cleaning
program treatment.  How can I clean used column?
Thanks!
Jimmy

__________________________________
Do you Yahoo!?
Exclusive Video Premiere - Britney Spears
http://launch.yahoo.com/promos/britneyspears/

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet