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CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: AXIAL RESOLUTION
Citation:
Zucker, R M. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: AXIAL RESOLUTION. MICROSCOPY TODAY 38:38-40, (2004).
Impact/Purpose:
To replace the subjectivity in evaluating a confocal microscope
Description:
Abstract
Confocal Microscopy System Performance: Axial resolution.
Robert M. Zucker, PhD
Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711
The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. When tests are made to evaluate the performance of a CLSM, the usual subjective assessment is accomplished by using a histological test slide to create a "pretty picture". Without the use of functional tests many of the machines may be working at sub optimal performance levels delivering sub optimum performance and possibly misleading data. In order to replace the subjectivity in evaluating a confocal microscope, tests were derived or perfected that measure field illumination, lens clarity, laser power, laser stability, dichroic functionality, spectral registration, axial resolution, scanning stability, PMT quality, overall machine stability, and system noise. It is anticipated by using this type of test data, performance standards for confocal microscopes will be obtained and the current subjectivity in evaluating CLSM performance will be eliminated. These tests will help serve as guidelines for other investigators to assess both the performance of their machines and the quality of data derived from their machines. Utilization of this proposed testing approach will help eliminate the subjective nature of assessing the CLSM and may allow different machines to be compared. These tests are essential if one is to make intensity measurements. These tests have been used in a similar manner to evaluate the performance of a Zeiss 510 Zeiss 510 Meta and a Leica AOBS confocal microscopy systems.