RE: cell culture for IHC
Luis,
The chambered slides we use are from Nalge Nunc, and are called Lab-Tek II
Chamber Slide System. We use the 4 chamber slide, with the "removable
chambers" , Nalge Nunc catalogue #154526. We buy them through VWR. They are
also available with 1,2 or 8 chambers.
Mary Georger
Center for Cardiovascular Research
University of Rochester Medical Center
KMRB Room 2-9816
601 Elmwood Avenue.
PO Box 679
Rochester, New York 14642
585-273-1548
> ----------
> From: Luis Chiriboga
> Sent: Friday, February 14, 2003 7:16 AM
> To: Chris van der Loos; histonet@pathology.swmed.edu
> Cc: bliven.laura@marshfieldclinic.org
> Subject: RE: cell culture for IHC
>
> Hi All
> I have seen a system that allows you to grow cells directly on glass
> slides
> . I do not know the name of the system since I have never actually used
> for cell culture but do have a collaborator who use it exclusively.
> Essentially, It is a clear plastic box without bottom that fits over the
> glass slide (excluding the label area). A "rubber?" contact adhesive is
> used(do not know name or composition) to attach the box to the slide
> surface
> such that a water tight seal is made. The box comes with a lid and the
> cells
> can be grown directly on the glass slide. The benefits of this system are
> that any fixative can be used. Also, if you are doing manual staining,
> the adhesive acts as a hydrophobic barrier so you do not have to use a PAP
> pen. The main drawback is if your are doing automated staining. You have
> to scrape the adhesive off the slide for cap gap instruments or ventana
> instruments because it will impeded the movement/mixing of solutions. In
> addition, the entire slide is covered with cells so it would require a
> lot
> of reagent when doing manual staining, we have gone around this by wiping
> of
> areas of the slide Ill see if I can come up with a name.
> Thought this might be another option (although the lack of vendor
> information makes it this post quite useless :-)
> Regards
> Luis
>
> -----Original Message-----
> From: Chris van der Loos [mailto:c.m.vanderloos@amc.uva.nl]
> Sent: Friday, February 14, 2003 2:53 AM
> To: histonet@pathology.swmed.edu
> Cc: bliven.laura@marshfieldclinic.org
> Subject: RE: cell culture for IHC
>
>
> Hi Laura,
> In the past I did some IHC on cell cultures grown on plastic disposables.
> This created an extra problem with respect to the fixation since
> acetone-fixation was not possible for obvious reasons. Finally, we used a
> 4% paraformaldehyde (freshly prepared in PBS) for 5 min at RT. This short
> aldehyde-fixation does not cause any cross-linking and therefore antigen
> retrieval is not needed (impossible too, because of the plastics!)
> During ALL IHC incubation steps and washings (from endo PO blocking up to
> chromogen) we included 0.1% saponin to open up the cell membranes (to get
> your IHC reagents in and out of the cells).
> Lots of success!
>
> Chris van der Loos
> Dept. of Cardiovascular Pathology
> Academic Medical Center
> Amsterdam, Netherlands
>
> Laura wrote:
> >Date: 13 Feb 2003 10:30:26 -0600
> >From: bliven.laura@marshfieldclinic.org
> >Subject: Cell Culture for IHC
> >
> >I have been asked to do an immunohistochemistry stain (West Nile Virus)
> on cell
> >culture. I usually on worked with paraffin blocks and smears. I'm not
> even
> >sure of the questions to ask, but here it goes....
> >In growing the cells, can I used glass slides (I don't have any
> plastic)?
> >Are the glass slides plain or charged?
> >How do I fix the cells? (I know if they're not fixed in formalin there
> will
> >be no antigen retrieval.)
> >Anything special I need to know before the jump such as potential
> problems or
> >background staining?
> >Thanks in advance,
> >Laura Bliven
> >Marshfield Laboratories
>
> Kathleen wrote:
> >Date: 13 Feb 2003 13:01:22 -0600
> >From: Kathleen Spencer
> >Subject: Re: Cell Culture for IHC
> >In the past I did IHC on cultured hippocampal neurons grown on round
> >glass coverslips in tiny petri dishes. The coverslips were coated with
> >poly-l-lysine after soaking in nitric acid. Everything was done with
> >sterile technique of course. The fixation of the cells and the staining
> >took place in the petri dish, suctioning off each solution. The
> >coverslips were placed cell side down on a slide with vectashield for
> >microsopy.
> >Hope this helps,
> >Kathleen
>
>
>
>
>
<< Previous Message | Next Message >>