Gene Expression: Confocal Microscopy

Baylor College of Medicine Intellectual and Developmental Disabilities Center
Center URL: https://www.bcm.edu/research/centers/iddrc/
Core URL: http://mrrc.bcm.tmc.edu/cores/confocal.html

Core Personnel
Core Director: Hugo Bellen, Ph.D., D.V.M.
Core Staff: Richard Atkinson, Ph.D.

Core Keywords
Gene Expression; Confocal Microscopy; Zeiss 510 systems

Confocal microscopy extends the capability of conventional fluorescence microscopy by enabling the investigator to visualize a thin optical plane within a sample. Out of focus portions of the sample are minimized in the image produced. Optical sections obtained confocally are superior to those obtained by mechanical sectioning, because consecutive sections remain in precise register and can be used to produce an accurate three dimensional image or model of the original sample.

Two Zeiss 510 systems (one upright the other inverted) are available for use by members of the MRDDRC. These newer systems incorporate many refinements and advances that improve the basic functions of older confocal systems and extend their usefulness for biological research. For example, the x,y,z stages allow automated collection of high resolution montage images of samples far larger than the field of view of a particular lens Improved automation of many collection and measurement functions both simplify use and improve mechanical precision of the systems. Time lapse and bleaching capabilities allow one to use recent methods such as Fluorescence Recovery after Photobleaching to study properties of motility of molecules within membranes and Fluorescence Resonance Energy Transfer to determine the extent of separation of molecules in the nanometer range. Live cell imaging is possible through the use of several specialized chambers for cells or tissue that permit heating and perfusion of a sample. Each microscope has an independent file server and an imaging workstation with software for preparation of publication quality figures.

Use of these systems by MRDDRC investigators has facilitated discovery and elucidation of genes involved in human disorders such as Rett and SCA1; functional analyses of genes including Math1, Mecp2, Ataxin, Gfi-1, shar-pei, VAP-33A, endophilin, crumbs, hrs, CSP, atonal, syntaxin, senseless, neurexin, ROP, discs lost, skittles, gutfeeling, synaptobrevin, synaptotagmin, apterous, Dpp and protein structure/function studies of potassium channel domains.

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Last Edited: 04/03/06 12:00 AM by Evette Mezger