Evaluation of Confocal Microscopy System Performance
| Abstract |
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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 could be working at suboptimal performance levels,
delivering suboptimum performance and possibly misleading data. 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, photomultiplier tube quality, overall machine stability, and
system noise. These tests will help serve as a guide for other investigators to ensure that their machines are working correctly
to provide data that are accurate with the necessary resolution, sensitivity, and precision. Utilization of this proposed
testing approach will help eliminate the subjective nature of assessing the CLSM and allow different machines to be compared.
These tests are essential if one is to make intensity measurements.
Affiliation(s): (2) Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC
Book Title: Cell Imaging Techniques: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 319 | Pub. Date: Nov-01-2005 | Page Range: 77-135 | DOI: 10.1007/978-1-59259-993-6_5
Subject: Imaging/Radiology
Key Words: Confocal microscope - lasers - coefficient of variation - photomultiplier tubes - field illumination - axial resolution - spectral registration - laser stability - beads - microscope lenses - quality assurance - quantification - spectroscopy
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