imaginal disc in situs

Jim Mahaffey jim_mahaffey at ncsu.edu
Fri Jun 8 17:01:25 EST 2001


Hi Mary,

I can't help with the background problem, but can offer a solution to the
tissue loss.  When doing antibody staining to imaginal discs, we leave the
discs attached to the inverted head. We just fix the whole inverted head
after removing much of the fat body etc.  That way, the discs are not loose.
After staining, we remove the discs and place them on slides to view.

Hope this helps,

Jim Mahaffey


"Mary Stewart" <Mary_Stewart at ndsu.nodak.edu> wrote in message
news:5.1.0.14.0.20010607180053.00aa0c60 at imap.ndsu.nodak.edu...
> Hi,
> Can anyone help us with advice on in situs to imaginal discs?  We made
> dig-labeled sense and antisense probes which worked great on embryos---no
> background with the sense probe.  We also have anti-dig antibody that was
> preabsorbed to embryos and this worked great on embryos.  When we tried
> these reagents on imaginal discs, we have a low, but unacceptable, level
of
> background with the sense probe.  The antisense gives a stronger signal,
> but we're not comfortable with the results since we have some overall
> general staining with the sense probe as well as some staining that
> resembles what we see with the antisense (even though the antisense
> staining is more intense).  Does anyone have some advice on how we might
> solve these problems?  Also, any advice on how to get through all the
> handling steps and minimize the amount of tissue we lose?  We lost a lot
of
> discs during all the handling and washing steps.  Thanks for any help.
> Mary Stewart
>
>
****************************************************************************
**
> Mary Stewart
> Assistant Professor
> Department of Biological Sciences
> North Dakota State University
> Office:  701-231-8226
> Lab:  701-231-6523
>





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