[Histonet] Rubber cement (for in situ)

[Bonnie J. McMahill]

We use the smaller coverslips (we have a large supply of 18 mm round ones that work really well), and apply the rubber cement around the edge with a 5 ml syringe.  It works very well and comes off easily after hybridization with a tweezer.

Bonnie McMahill
Histology Supervisor
InCyte Pathology
Spokane, WA


> -----Original Message-----
> From:	Connolly, Brett M [SMTP:brett_connolly <@t> merck.com]
> Sent:	Thursday, February 03, 2005 9:46 AM
> To:	'- -'; histonet <@t> lists.utsouthwestern.edu
> Subject:	RE: [Histonet] Rubber cement (for in situ)
> 
> Don't use the brush. Use 22 mm wide coverslips instead of 24 mm. With a
> plastic transfer pipet lay a thin line of rubber cement at the junction of
> the coverslip and the slide. After hybridization the solidified rubber
> cement easily peels away. Actually, your temps are relatively low for ISH,
> we've hybridized at >65C. 
> 
> Brett
> 
> Brett M. Connolly, Ph.D.
> Merck & Co., Inc.
> MRL, Imaging Research
> WP26A-3000
> PO Box 4
> West Point, PA 19486
> PH 215-652-2501
> fax. 215-652-2075
> e-mail. brett_connolly <@t> merck.com
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of - -
> Sent: Thursday, February 03, 2005 12:21 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Rubber cement (for in situ)
> 
> 
> Hello,
>  
> I am starting to work up a protocol for in situ hybridization and discovered
> I need to seal my slides because of the high temps and long incubation.  Can
> anyone help me with the following questions:  
>  
> a. I noticed the rubber cement is quite fluid.  Using the brush that it
> comes with the can (or another method), how does one go about applying it
> after your sample is coverslipped?
>  
> b. I've heard about fumes ... what concerns should I have; safety
> precautions should I take? (my normal range of temperature will be between
> 43 and 55 degrees Celcius within a Boekel Slide Moat or all purpose
> incubator)
>  
> c. Are there better methods to use when sealing slides? 
>  
> d. What is the best way to remove the rubber cement/coverslip after
> performing hybridization?
>  
> THANK YOU ALL VERY MUCH!!!!!
> 
> 		
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