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Microarray
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Overview
Integral to understanding the function of a gene product is the determination
of which cells it is in, where within those cells it is localized, and
what happens to those cells when the gene is mutated or deleted. These
analyses typically include microscopy. The Microscopy and Imaging Module
will provide a microscopy core that will interface with the other modules
by assisting investigators in such analyses. Specifically, it will provide
assistance with tissue processing, sectioning, histochemistry and immunoloabeling
(including immunofluorescence), in situ hybridization, light and electron
microscopy, and digital imaging procedures. It will provide some relief
for routine work for those labs that are already accomplished in microscopy,
and it will provide technical assistance for those labs that are not.
Resources
This core module is staffed by two experienced Research Technicians whose
roles is to: (1) train individuals in the use of the core microscopy equipment,
and oversee the use and care of the equipment; (2) perform immunolabeling
and in situ hybridization procedures, including sectioning. Procedures
will include processing, sectioning, and staining and immunolabeling of
8-10 mm frozen sections, semi-thin plastic (e.g. Epon, L.R. White) sections,
and ultrathin sections, depending on the level of resolution required.
All these procedures will be performed in the core module facility, as
will all analyses at the light level. Analysis of electron microscopy
specimens will be carried out on electron microscopes that are available
on a user fee basis in facilities either at The Scripps Research Institute
or UCSD. Semithin or ultrathin cryosections may also be used in immunolabeling
studies; these sections would be obtained with cryoultramicrotomes available
on a user fee basis in these same EM facilities, or on a cryoultramicrotome
in Dr Williams' lab.
The Research Technicians will also assist in the analysis of specimens.
Light microscope images will be acquired directly in digital form, using
a Zeiss Axioplan II microscope equipped with a Photometrics Quantix CCD
camera. Digital EM images will obtained by scanning negatives generated
by the electron microscopes. The Research Technician will also be responsible
for assisting with basic image processing in Adobe Photoshop (including
optimizing signal-to-noise, image merging, and color balancing) and will
provide prints of selected images to investigators as well as CDs or Zip
disks with the raw images for storage and/or further processing and analysis.
The computer-controlled Zeiss microscope and digital camera system can
acquire high quality light microscopical images. Furthermore, digital
image acquisition with a high resolution, sensitive CCD camera is available
for immunofluorescence imaging of the low abundance. In some cases, such
as in the analysis of the structure of the photoreceptor connecting cilium
and the localization of proteins, it will be necessary to use electron
microscopy. A negative scanner that can accommodate the large-format,
high resolution EM negatives enables investigators to bring back EM images
into the module for analysis in the same way as the light microscopy images.
Resources available
This Microscopy and Imaging Module has a Reichert-Jung Cryostat for thick
frozen sections of eye tissues, A Zeiss Axioplan II microscope equipped
with phase, DIC, and epifluorescence optics, and a Photometrics Quantix
CCD camera interfaced with OpenLab software on a Macintosh G4 computer
to obtain high quality digital images of the multiple specimens. More
recently, a BioRad MRC1024 scanning laser confocal microscope and a spinning
laser disc attachment for the Zeiss Axioplan have been acquired and are
available to Core group members.
For information regarding this site, contact Suzanne Bacon: bacons@scripps.edu
© 2003 The Scripps Research Institute - All Rights Reserved