I used to do it by having all the stains and reagents in a series of
watch-glasses (any shallow dish would do) and transferring each section in
turn from floating on the liquid in one dish to the next using a "hockey
stick" - a piece of thin glass rod with one end bent at an angle in a Bunsen
flame, so that it looks like an ice-hockey stick rather than a field-hockey
stick. One soon learns how to pick up the sections without letting them
stick to themselves and how to put them into the next dish by rolling them
off the hockey stick from one side. At the end you put them onto slides the
same way and mount in aqueous mountant. If you want to dehydrate and clear,
do it before mounting, and use DPX or whatever. As far as I can remember (it
was a long time ago), I used normal stains for normal times. Are the
sections in celloidin, by any chance? If they are, you still treat them as
above, but then you will need more dilute stains for longer times.

Lesley Weston.


on 29/10/2002 11:27 AM, Greg Dobbin at dobbin@Upei.CA wrote:

> HI Melissa,
> I don't have a lot of experience with this, but what I did do for
> someone else one time, was to cut 50-80 um frozen sections and
> place them directly in a 24-well plate (each section to its own well).
> The 24-well plate was kept in the cryostat and the fixative (I forget
> now, which one they were using) was already dispensed to all the
> wells. All subsequent staing took place in the well to minimize
> tissue handling (ie the reagents were changed rather than moving
> the tissue sections). I wasn't present to see the sections get
> mounted on the slide, but I understand it was a delicate process!
> Good luck.
> 
> 
> Date sent:          Tue, 29 Oct 2002 11:39:17 -0800 (PST)
> From:               Melissa Jans 
> Subject:            staining of free floating sections
> To:                 histonet@pathology.swmed.edu
> 
>> 
>> --Boundary_(ID_y10JQ1AS4sM3MYrPcmOmxA)
>> Content-type: text/plain; charset=us-ascii
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>> 
>> 
>> I have some 100, 200 and 400 micron thick lung sections that a pathologist
>> would like to have stained.  These sections are loose (not adhered to a
>> slide) and they do not have paraffin in them (don't ask me how they did this
>> but they did).  He would like me to do an H&E, Alcian Blue/PAS,
> Trichrome, and and Elastichrome.  Does anyone have experience doing this?
> What is the best way to transfer your loose section from reagent to reagent?
> Do I need to dilute my solutions and/or change my staining times?
>> 
>> Any help would be appreciated, this is something totally new to me.
>> 
>> Thanks!!
>> 
>> Melissa Jans
>> 
>> Lead Scientist, Histology Lab
>> 
>> University of Iowa Healthcare
>> 
>> 
>> 
>> ---------------------------------
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>> 
>> --Boundary_(ID_y10JQ1AS4sM3MYrPcmOmxA)
>> Content-type: text/html; charset=us-ascii
>> Content-transfer-encoding: 7BIT
>> 
>> 
I have some 100, 200 and 400 micron thick lung sections that a pathologist
>> would like to have
> stained.  These sections are loose (not adhered to a slide) and they do
> not have paraffin in t
> hem (don't ask me how they did this but they did).  He would like me to
> do an H&E, Alcian
> Blue/PAS, Trichrome, and and Elastichrome.  Does anyone have experience
> doing this?  What
> is the best way to transfer your loose section from reagent to reagent? 
> Do I need to dilute
> my solutions and/or change my staining times?  
>> 
Any help would be appreciated, this is something totally new to me.
>> 
Thanks!!
 
>> 
Melissa Jans
>> 
Lead Scientist, Histology Lab
>> 
University of Iowa Healthcare
 

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