[Histonet] Mouse Embryo Processing

[Stephanie Schmoll]

Sorry I'm new at the histonet!
I am using a Leica ASP 300 tissue processor.  All of the reagents run 
at room temp and paraffin is at 57C suitable for the Tissue Prep2 
paraffin I am using.  I am embedding whole embryos developmental days 7 
through 18 and neonate.  I am only having problems with the late stages.


On Tuesday, October 14, 2003, at 05:32 PM, Pamela Marcum wrote:

> Stephanie,
>
> Are these whole embryos?  Do they have all of the skin in place?  Can 
> you
> give a size estimate?  Are you using a tissue processor and if so 
> where are
> you using heat (other than paraffin) and vacuum?  What kind of tissue
> processor?  Please give temperatures also.  Thanks,  Pam Marcum
>
>> -----Original Message-----
>> From: histonet-admin <@t> lists.utsouthwestern.edu
>> [mailto:histonet-admin <@t> lists.utsouthwestern.edu]On Behalf Of Stephanie
>> Schmoll
>> Sent: Tuesday, October 14, 2003 4:53 PM
>> To: histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] Mouse Embryo Processing
>>
>>
>> I am fairly new to tissue processing with paraffin, have always done
>> cryo-embedding in the past.  I am having trouble processing late stage
>> mouse embryos, day 17 and 18.  The paraffin doesn't seem to be
>> infiltrating.  Any suggestions would be appreciated.
>>
>> My current protocol is as follows:  24hrs 10% NBF, 70% EtOH 30 min, 
>> 80%
>> 30 min, 95% 30 min x2, 100% 30 min x2, Xylene 30 min x2, paraffin 30
>> min x3.
>>
>>
>> Stephanie Schmoll, Morphology Core Manager
>> Center for Organogenesis
>> University of Michigan Medical School
>> 5795C  Medical Science Bldg.  II
>> Phone: (734) 647-9021
>> Fax: (734) 647-9559
>>
>>
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>>
>
>
Stephanie Schmoll, Morphology Core Manager
Center for Organogenesis
University of Michigan Medical School
5795C  Medical Science Bldg.  II
Phone: (734) 647-9021
Fax: (734) 647-9559