The Leica TCS-NT confocal microscope has been installed in Room 006 of MSRB. This microscope forms the heart of the UNC Neuroscience Center Confocal Facility, so named because the microscope was purchased with funds obtained through the Howard Hughes Research Resources for Medical Schools grant, awarded to strengthen UNC's neuroscience effort. Physiology Department members not in the Neurobiology Program will be welcome to use it since our Department contributes space, financing, and expertise to the instrument and its setup.

Confocal microscopes are used mainly for fluorescence. Relative to a conventional microscope, they increase resolution and contrast by illuminating only a tiny portion of the field of view at any instant and by removing from the image that light emitted from specimen planes above and below the plane of focus. This amounts to "optical sectioning". The entire image field is examined by scanning the laser beam across the sample, hence the name Confocal Laser Scanning Microscopy. Because of the optical sectioning capability, the confocal gives XY-plane images of living or fixed specimens with high resolution and little glare. Volume images can be gathered by scanning in the X-Y plane at multiple Z levels. These images can then be reconstructed as 3D renders of the sample. Other modes of operation such as timed captures which may be assembled into time-lapse movies are possible.

The Leica is presently the top of the heap in confocal microscopes of its type. It is built around a Leica inverted microscope, a PC Pentium Pro 200MHz computer with Windows NT 4.0, and three lasers. The lasers emit visible light, so while we won't have UV capability, we will have terrific triple-label capability using FITC/BODIPY/CY2, Rhodamine/Texas Red/CY3, and CY5 or similar fluorophores. Differential interference contrast images can be made at the same time as the fluorescence image.

Since installation, we have been learning the ins and outs of the instrument and developing a teaching approach which we are testing on willing victims. Included in the teaching approach is a guide written at a very practical level which will be available on paper for thorough study and on the World Wide Web for quick consultation. It is our aim here to be able to turn users loose on the microscope with complete confidence that it will be used properly and efficiently, and that the next user will be fully respected. Being well prepared in these areas will be of major importance once the entire molecular and cellular neuroscience community is invited to use the microscope.

The fun but challenging aspects of confocal microscopy include optics, computing and digital analysis of images. This is all summarized in several inches of textbook you can consult when our guide proves insufficient. For the computing aspects of all this, M. C. is right in his element, and we anticipate offering fine instruction in the computer aspects of the microscope and image analysis as well as image acquisition.

					© 1998-2003 Michael Chua, Cell & Molecular Physiology, UNC.  All rights reserved.
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