The Confocal Imaging Facility (CIF) of the NIA IRP has been in operation since October 2004. This facility has one older model (Zeiss LSM 410) confocal system with low usage by IRP investigators. The NIA CIF has been managed and overseen with the expansion and successfully integration of a state of the art Zeiss LSM 510 Meta confocal system, updating the Zeiss Axioskop fluorescent microscope MCID/QImage analysis system and using Metamorph/Coolsnap imaging and analysis system. This has resulted in extensive collaboration with intramural scientists at different levels of expertise ranging from principal senior investigators through postdoctoral fellows and intramural research trainees. We have trained over thirty investigators to be independent users of the confocal microscope and have held scores of consultations with NIA researchers to assist with their imaging and image analysis. We have helped users integrate imaging into their experimental systems and in troubleshooting problems. This has resulted in an increase in the use of immunofluorescence and imaging at the IRP, as attested by the widespread use of the CIF last year by over 60 researchers, ranging from post-bacs to senior investigators.? ? The following techniques have been used or introduced at the CIF: (a) Use of confocal microscopy for precise sub-cellular localization and co-localization of proteins; (b) Advanced immunological, biochemical and imaging techniques required for the investigation of intracellular and intranuclear protein trafficking; (c) Development of methodology to cause DNA damage to live cell DNA at the sub-micron level by use of the continuous scanning UV laser of the LSM 510 Meta system; (d)Time-lapse, FRAP and ratio-metric analysis of cellular processes in live cells using the 510 Meta confocal microscope; (e) Volumetric (3D) reconstruction of intracellular protein distribution using confocal or deconvolution techniques.
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