[Histonet] IHC withquantum dots

[Richard Cartun]

Have you tried staining a section of tonsil to see if you get the same pattern of staining.  I have found "precipitation" to be a problem with alkaline phosphatase (PA) detection, although I am by no means an expert when it comes to AP IHC.  I use HRP/AEC for most of my infectious disease studies including Bartonella henselae.  Don't forget that you may not see the normal morphology of the bacterium with IHC.  You are labeling epitopes on the surface of the organism; you are not staining the entire cell wall.  I have found B. henselae to be one of the most difficult organisms to label (and interpret) with IHC.  Good luck!

Richard 


Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

>>> "Joanne Mauger" <MAUGER <@t> email.chop.edu> 10/15/04 03:45PM >>>
Hi group,
Can anyone help me with suggestions-I am working up the monoclonal Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have a purchased control from Newcomer Supply. I am using permanent red chromagen from Dako., with alk phos link also from Dako. I am getting a nice specific staining of what looks like specs vs tiny rods or spirochetes, but the control tissue does , by Warthin Starry, look much more spirochete-ish. Can I trust this result? I have also stained several suspected + cases and see the same dust like + staining,but I am afraid it is teeny bits of precipitate. 
Does this ring a bell?
Please help.
Thanks,
Jo

>>> "John Kiernan" <jkiernan <@t> uwo.ca> 10/13/04 09:57PM >>>
Quantum dots are nanocrystals of a semiconductor
(exact chemical identity seems to be trade secret).
They may have a future in a new generation of
tiny computers.

Quantum dots fluoresce in a wide range of colours 
and can be conjugated to antibodies, avidin,
phalloidin etc. You can buy conjugated reagents
or kits to conjugate your own.
The Quantum Dot Corporation's web site
http://www.qdots.com/live/index.asp 
shows some pretty pictures of labelled cells.

There is a recent paper about quantum dots in the
J. Histochem. Cytochem. Here's the bibliographic
information and abstract, from
http://www.jhc.org/cgi/content/abstract/52/1/13 
If your institution subscribes to the journal you'll
be able to get the full text on line.

---------
Rozalia Nisman, Graham Dellaire, Ying Ren, Ren Li and David P.
Bazett*Jones
(2004)
Application of Quantum Dots as Probes for Correlative Fluorescence,
Conventional, and Energy-filtered Transmission Electron Microscopy 
J Histochem Cystochem 52:13*18

Luminescent semiconductor quantum dots (QDs) are a new class of
fluorescent label with wide-ranging applications for cell imaging. The
electron density and elemental composition of these materials permit the
extension of their use as probes in conventional electron microscopy
(TEM) and energy-filtered TEM (EFTEM). Here we illustrate the
feasibility of using streptavidin-conjugated QDs as TEM tags by labeling
a nuclear protein on cell sections and obtaining correlative
fluorescence and TEM data. We also show that QD probes can be employed
in conjunction with immunogold for co-localization of proteins at the
ultrastructural level. Furthermore, by obtaining cadmium elemental maps
of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the
potential of QDs for co-localization of multiple proteins when used in
combination with EFTEM. 
---------

A Google search for quantum dots brings up thousands of hits.
Searching for "quantum dots" and histochemistry and cytochemistry
brought up 60 hits, at least half of them in English and
relevant.

John Kiernan
London, Canada.
-----------------------------------------------
Patsy Ruegg wrote:
> 
> What are "quantum dots"?
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu 
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anna
> Elisse Beaudin
> Sent: Wednesday, October 13, 2004 7:55 AM
> To: histonet <@t> lists.utsouthwestern.edu 
> Subject: [Histonet] IHC withquantum dots
> 
> Hello,
> 
>   Has anyone ever done immunohistochemistry on tissue sections using
> primary (or secondary) antibodies conjugated to quantum dots?  if so, do
> you think it would be possible to do multiple labeling using different
> colored quantum dots?  thanks in advance for your help!
> 
> Anna Beaudin
> Division of Nutritional Sciences
> Cornell University
-------------------------------------------------------

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