Quality assessment of confocal microscopy slide based systems: performance.
BACKGROUND
All fluorescence slide-based cytometry detections systems basically include the following components: (1) an excitation light source, (2) intermediate optics, and (3) a detection device consisting of a CCD camera or a PMT. The optical principles employed is slide-based systems are similar to those of confocal microscopes (CLSM).


METHODS
The following tests evaluated confocal equipment performance: dichroic reflectivity, field illumination, lens performance, laser power output, spectral registration, axial resolution, PMT reliability, and system noise.


RESULTS
Quality assurance tests provide a basis to determine if the equipment is operating correctly. Laser power, PMTs function, dichroic reflection, spectral registration, axial registration, system noise and sensitivity, lens performance and laser stability were tested colocalization of UV and visible peaks of a bead should be less than 210 nm. Interference contrast optics decrease fluorescence resolution.


CONCLUSIONS
QA tests that assess CLSM system performance are also applicable to other slide-based systems. By utilization this type of testing approach, the subjective nature of assessing the CLSM may be eliminated. These tests serve as guidelines for other investigators to ensure that their machines are providing data that is accurate with the necessary resolution, sensitivity and precision.
Zucker
Robert M
RM
Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Researchand Development US Environmental Protection Agency, Research Triangle Park, NC 27711, USA. zucker.robert@.epa.gov
D000818:Animals; D002645:Chickens; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods* / Q000592:standards*; D012015:Reference Standards