[Zbrafish] Re: Bleaching embryos after In Situ Hybridization

[Qiyan Mao]

Hi,

I've recently bleached my 48 h embryos before ISH according to Thisse's
protocol (Nature protocols (2008) 3(1):59) using 3% hydrogen peroxide in
0.5% KOH, and it worked fine for me. The whole bleaching takes around 20
minutes. My impression is that this bleaching step doesn't alter ISH
staining. I also tried post-ISH bleaching using the same procedure, but for
some reason the pigment doesn't go away as easily.

Hope this could help.

Best, Qiyan


On 8/20/08 12:03 PM, zbrafish-request from oat.bio.indiana.edu wrote:

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>    1. Re: Bleaching embryos after In Situ Hybridization
>       (nonolow from gmail.com)
>    2. Re: Bleaching embryos after In Situ Hybridization
>       (nonolow from gmail.com)
> 
> 
> ----------------------------------------------------------------------
> 
> Message: 1
> Date: Wed, 20 Aug 2008 09:49:09 -0700 (PDT)
> From: nonolow from gmail.com
> Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization
> To: bionet-organisms-zebrafish from moderators.isc.org
> Message-ID:
> <69b4622d-afa1-4d79-bcf1-d140589920dc from f36g2000hsa.googlegroups.com>
> Content-Type: text/plain; charset=ISO-8859-1
> 
> bleaching is a routine approach in frog embryo whole mount in situ
> hybridization, i think you could find the protocal and recipes in the
> Frog Book (Early Development of Xenopus Laevis: A Laboratory
> Manual)....
> And be aware the time of bleaching, hydrogen peroxide could breakdown
> the staining of in situ hybridization.
> 
> 
> low
> 
> 
> 
> On Aug 19, 1:22 am, shellyleib... from gmail.com wrote:
>> Hello
>> I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I
>> wanted to ask if anyone knows how to bleach the pigment of 2dpf
>> zebrafish embryos post whole mount In Situ Hybridization (it was not
>> raised in PTU water).
>> Many thanks in advance
>> 
>> Shelly
> 
> 
> 
> 
> ------------------------------
> 
> Message: 2
> Date: Wed, 20 Aug 2008 09:55:33 -0700 (PDT)
> From: nonolow from gmail.com
> Subject: [Zbrafish] Re: Bleaching embryos after In Situ Hybridization
> To: bionet-organisms-zebrafish from moderators.isc.org
> Message-ID:
> <59eb3d60-d642-4604-87fc-52e9cd3dd91d from z72g2000hsb.googlegroups.com>
> Content-Type: text/plain; charset=ISO-8859-1
> 
> bleaching is a routine approach in frog embryos, you could find some
> protocols and recipes in the book, Early Development of Xenopus
> Laevis: A Laboratory Manual,
> be aware the time of bleaching, hydrogen peroxide could breakdown the
> staining of in situ hybridization.
> good luck
> 
> 
> low
> 
> 
> 
> 
> 
> On Aug 19, 1:22 am, shellyleib... from gmail.com wrote:
> 
> 
>> Hello
>> I'm Shelly, a PhD student at Gothilf's lab from Tel Aviv University. I
>> wanted to ask if anyone knows how to bleach the pigment of 2dpf
>> zebrafish embryos post whole mount In Situ Hybridization (it was not
>> raised in PTU water).
>> Many thanks in advance
>> 
>> Shelly
> 
> 
> 
> 
> ------------------------------
> 
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> End of Zbrafish Digest, Vol 39, Issue 10
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