Integrated Microscopy Shared Resource
(Michael A. Mancini, Ph.D., Director)
The goal of the Integrated Microscopy shared resource is to provide state-of-the-art light and electron microscopic imaging, image analysis resources, and cellular-image-based assay development assistance to researchers in the Dan L. Duncan Cancer Center, at Baylor College of Medicine, and throughout the Texas Medical Center.
The Integrated Microscopy shared resource houses state-of-the-art instruments for digital transmitted light microscopy, epifluorescent microscopy, and transmission electron microscopy. Transmitted light microscopy capabilities include phase contrast and differential interference contrast . The epifluorescent microscopes offer standard widefield, confocal, and deconvolution capabilities. All of the above techniques are compatible with live-cell time-lapse imaging, automated multipoint imaging with autofocusing, as well as with multi-well plate specimen formats. Two high-throughput widefield epifluorescent microscopes are available for fast, automated image acquisition of multiwell plates. Our confocal microscopes enable FRAP/FLIP and other laser-based experimental techniques. A high resolution digital TEM is available and TEM specimen preparation service is offered. The Integrated Microscopy shared resource also provides access to numerous image analysis software resources.
Day-to-day operations are managed by Debra Townley, B.A, and James Broughman, Ph.D, who have extensive training and experience in electron and light microscopic methods. To ensure that the goals of individual investigators are reached, experimental design and interpretation consultation are available with the Integrated Microscopy shared resource director, Michael Mancini, Ph.D. Moreover, the facility provides extensive instrument training for new users and more advanced and customized training upon request.
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 DLDCC 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, DLDCC principle investigators are required to submit a one page proposal describing the goals of the assay to Dr. Mancini (by e-mail). An ad hoc panel will assess proposals for compatibility/feasibility with the current resources.
The following assays are currently 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.
- Hitachi H7500 TEM
For ultra-high magnification examination of thinly sliced cells, internal cellular structures and/or organelles. High resolution digital images (2048x2048) are obtained by use of the built-in Gatan US1000 camera.
- TEM Specimen Sectioning
Thick-sections (500nm – 995nm) can be cut and stained for light microscopy by a trained technician. Thin-sections (70nm-80nm) for examination on the Hitachi H7500 are also produced upon request.
- TEM Specimen Staining
Special staining techniques are used to enhance contrast and reveal more distinct ultra-structure when viewing thin-sections on the Hitachi H7500.
- TEM Tissue Processing
Tissue can be processed according to a specific protocol provided by the Investigator, or according to a globally accepted standard protocol.
- MT6000 Ultramicrotome
This ultramicrotome is provided for those who prefer to do their own sectioning for TEM, or for those who wish to learn.
- DeltaVision Deconvolution Microscope
High-precision z-drive enables three dimensional image acquisition and restorative image deconvolution for the production of extremely high quality three dimensional images free of out of focus light.
DeltaVision Core Deconvolution Microscope
As above, plus heat/humidity/CO2 controlled live cell chamber; autofocus; multi-well compatible; multipoint and time-lapse capability; highly sensitive and low noise Cascade II EMCCD camera for very fast image acquisition; multiplex mode for near-instantaneous filter switching.
- Nikon A1Rs
The Nikon A1Rs confocal laser scanning microscope has both high resolution (4Kx4K) and high speed (up to 230 frames/sec) imaging capabilities. The system can perform real time RFAP, calcium imaging (with appropriate dyes), FRET, and time lapse studies of live cells.
- Beckman Coulter IC-100 High-Throughput Microscopes
Automated widefield epifluorescent imaging of entire multi-well plates in up to four fluorescent channels.
- SAMI and BioMek NX Plate Processing Robot
Automated multi-well plate cell seeding and specimen staining.
- Image Analysis Software Workstations
Pipeline Pilot, CyteSeer, Imaris, MetaMorph, SoftWorx (Applied Precision), Image Browsers (Zeiss, Applied Precision).
- Assay Development Support
Full range of state-of-the-art “high content analysis” including nuclear translocation, reporter gene analysis, cell cycle, apoptosis, etc.
Charge Back Fees and Forms
- View pricing for all services.
- Visit the Integrated Microscopy new user page to fill out an online new user request.
Associate Professor, Molecular and Cellular Biology
Director, Integrated Microscopy Shared Resource
James Broughman, Ph.D.
Shared Resource Manager
Cullen Building, Room 120A, 123A, 124A
Baylor College of Medicine
One Baylor Plaza
Houston, TX 77030
Please contact us through our Microscopy Page