DiI direct crystal placement tips

[matthewmakes]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

hi.

i've been trying to label small numbers of cells in the DM of chicken embryos.  i've been trying to use iontophoresis, but the technique makes me want to pull my hair out.  i was hoping for something a bit simpler.  i don't have a detailed protocol for placing DiI crystals.  all i can seem to find is, "pickup the crystal on the tip of a glass needle and place it on the tissue."  i've played with needle size and crystal size.  i seem to either get the crystal stuck in the needle, and then it doesn't do much labeling.  either that or the crystal comes off the tip of the needle when it hits the solution and floats.  i'd really appreciate any advice.

it'd also be nice if someone can suggest any good alternatives.  i'm trying to label small numbers of cells in the DM.  after giving time for the peeled back ectoderm to heal, cross section, counterstain, live imaging.

thank you,  matthew

---

Keep your kids safer online with Windows Live Family Safety. Help protect your kids.

[Eric Scarfone]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hey
Non-specific GFP transfection with a generic promoter will give you
random cell labelling.
I have used the gene-gun to that effect. It has a pretty low
transfection rate and you get a few labelled cells here and there
among mostly unlabelled ones.
I agree that DiI is a pain, When it works it is either too much or too
little!
Cheers
Eric


Eric Scarfone, PhD, CNRS,
Center for Hearing and communication Research
Department of Clinical Neuroscience
Karolinska Institutet

Postal Address:
CFH, M1:02
Karolinska Hospital,
SE-171 76 Stockholm, Sweden

Work:  [phone],
Cell:  [phone]
Fax:   [phone]

email:  [hidden email]
http://www.ki.se/cfh/


----- Original Message -----
From: Matthew Smith <[hidden email]>
Date: Wednesday, July 23, 2008 11:14 pm
Subject: DiI direct crystal placement tips
To: [hidden email]

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> hi.
>
> i've been trying to label small numbers of cells in the DM of
> chicken embryos.  i've been trying to use iontophoresis, but the
> technique makes me want to pull my hair out.  i was hoping for
> something a bit simpler.  i don't have a detailed protocol for
> placing DiI crystals.  all i can seem to find is, "pickup the
> crystal on the tip of a glass needle and place it on the tissue."  
> i've played with needle size and crystal size.  i seem to either
> get the crystal stuck in the needle, and then it doesn't do much
> labeling.  either that or the crystal comes off the tip of the
> needle when it hits the solution and floats.  i'd really
> appreciate any advice.
>
> it'd also be nice if someone can suggest any good alternatives.  
> i'm trying to label small numbers of cells in the DM.  after
> giving time for the peeled back ectoderm to heal, cross section,
> counterstain, live imaging.
>
> thank you,  matthew
>
> _________________________________________________________________
> Keep your kids safer online with Windows Live Family Safety.
> http://www.windowslive.com/family_safety/overview.html?

ocid=TXT_TAGLM_WL_family_safety_072008