Integrated Microscopy

(Michael A. Mancini, Ph.D., Director)

Dr. Michael Mancini For over 30 years, the Integrated Microscopy Core has provided state-of-the-art light and electron microscopy imaging support for dozens of laboratories throughout Baylor College of Medicine. This fully digital-imaging-based resource houses routine microscopy (Zeiss, CoolSnap CCD, Metamorph software), multispectral laser scanning confocal microscopy (Zeiss LSM 510 Meta), deconvolution-based microscopy (Zeiss, Applied Precision), a high resolution digital TEM (Hitachi 7500/Gatan 2x2K), and a fully automated high throughput micrscopy suite (Nikon, Beckman Coulter), including high throughput fluid handling robotics (Beckman Coulter).

Day-to-day operations are managed by a Jeannie Zhong, B.A., M.A. and Deborah Townley, B.A, two research associates with extensive microscopy and photography experience, with a third technician starting in June 06. To ensure that the goals of individual investigators are reached, experimental design and interpretation consultation are available with the Shared Resource Leader, moreover, the facility provides extensive instrument training for new users. Web-based communication facilitates core organization, data archiving and collaboration. Identification of labeled proteins, nucleic acids, organelles, and cell skeletons are now standard (e.g. antibodies and/or "GFP" technology). The facility supports live cell assays for time-lapse imaging, protein-protein interactions (FRET), molecular dynamics of complex formation/disassembly (FRAP/FLIP), and combined approaches utilizing creative stable cell lines that facilitate live read out of transcription/translation/chromatin modeling.

About 23 Cancer Center members use this facility and are charged a nominal fee for instrument use (e.g., $12.50-30/hr for use of the various microscopes). Equipment grants, strong ties with equipment and software manufacturers, and imaging-related biotech compains are used to maintain cutting-edge capabilities.

High Throughput Assay Development

With support from DLDCC resources, the Integrated Microscopy Shared Resource is proud to announce the establishment of a High Throughput Microscopy-based Assay Development Resource for all DDLCC investigators.

This new resource will provide consultation and technical support for pilot experiments using coverslips, or 96 or 384 well plate formats in single cell fluorescence-based imaging studies and quantitative image analyses. To take advantage of this new service, DDLCC principle investigators are required to submit a ONE PAGE proposal describing the goals of the assay to Dr. Mancini (by email). An ad hoc panel will assess proposals for compatibility/feasibility with the current resources. The following assays are currently already available or in development, with customization likely needed dependent upon cell lines, antibodies, fluorescent probes, reporters, etc.

Nuclear translocation assays (e.g., cytoplasmic protein trafficking to the nucleus)
Subnuclear pattern assays (e.g., intranuclear "speckling")
Cell proliferation and/or nuclear size and cell cycle assays (e.g., akin to DNA content-based assays using cell sorting)
Apoptosis assays (e.g., using DNA-based and/or apoptosis marker approaches)
Fluorescent transcription reporter gene assays (e.g., using fluorescent protein reporters)
Plasma membrane trafficking (e.g., cytoplasmic protein translocating to the plasma membrane).
Measurement of cytoplasmic 'foci' (e.g., mitochondria, lipid droplets, etc)

In conjunction with the John S. Dunn Gulf Coast Consortium for Chemical Genomics (GCC CG), several assays are now in development and/or in small scale screening mode. Access to the following GCC-CG libraries is available at an "at cost" basis for materials and machine time.

Human Kinase siRNA library (635, Invitrogen Stealth)
Human Nuclear Receptor siRNA library (281, Invitrogen Stealth)
Human Nuclear Receptor CoRegulator siRNA library (49, Invitrogen Stealth)
Chembridge “Discovery” small molecule collection (30,000)
Prestwick off-patent drug library (1,100)

Interested PIs should write Dr. Mancini directly to request a short questionnaire to facilitate project feasibility.

Fees:

The Integrated Microscopy Shared Resource fees are as follows:

Resource	non-CC rate	CC rate (20% discount)
Confocal Microscopy	35/hr	28/hr
Decovolution Microscopy	35/hr	28/hr
Axioplan 2	17.50/hr	14/hr
Digital TEM	35/hr	28/hr
Image Analysis Workstations	7/hr	5.60/hr
Ultramicrotome	7/hr	5.60/hr

Assay Development for High Throughput Microscopy

	non-CC rate	CC rate (50% discount)
Technical Support/Tutorials	35/hr	17.50/hr
High Throughput Microscopy	35/hr	17.50/hr
Image Analysis (Pipeline Pilot)	10/hr	5/hr
Microfluidic Robot (BioMek NX)	15/hr	7.5/hr
TitreTek robotic cell plating	10/run	5/run

All disposable items (plates, tips, etc) and reagents are to be provided by the CC Investigator.

Please contact us through our Microscopy Page