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Research Technologies Branch

Biological Imaging Section

Technology

Confocal Microscopy

Instruments:

Leica TCS-NT/SP1, Leica SP2—AOBS

Two confocal microscopes are available for analysis of fluorescent labeled samples. Instruments can be operated independently by users after completion of training, or can be operated by a member of the facility. In order to allow time for the installation of appropriate equipment, please note (when scheduling your time) whether live cell imaging is to be done.

Confocal microscopy is a specific form of fluorescence microscopy. Samples are labeled in the same manner as for routine epifluorescence microscopy. Confocal microscopy allows for greatly improved x, y, and z axis resolution and for imaging of dual and triple labeled samples without crosstalk. With high resolution in the z axis, optical sections can be collected free of out-of-focus fluorescence from adjacent layers. These optical sections can be stacked and projected to give 3D images of the sample. Time lapse confocal microscopy can provide 3D data through time to observe movement of or within cells. The principle differences between confocal and epifluorescence microscopy are the presence of the confocal pinhole and the substitution of lasers for the standard mercury lamp.


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  • Division of Intramural Research (DIR)