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Quality Assurance Testing for Modern Optical Imaging Systems

Published online by Cambridge University Press:  11 April 2011

Robert F. Stack ,
Carol J. Bayles ,
Anne-Marie Girard ,
Karen Martin ,
Cynthia Opansky ,
Katherine Schulz  and
Richard W. Cole
Robert F. Stack
Affiliation:
Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY 12201, USA
Carol J. Bayles
Affiliation:
Cornell University, Life Sciences Core Laboratory Center, Weill Hall, Ithaca, NY 14853, USA
Anne-Marie Girard
Affiliation:
Oregon State University, Center for Genome Research and Biocomputing, 3021 Ag Life Sciences Bldg., Corvallis, OR 97331, USA
Karen Martin
Affiliation:
West Virginia University, Department of Neurobiology and Anatomy, P.O. Box 9128, Morgantown, WV 26506, USA
Cynthia Opansky
Affiliation:
Blood Center of Wisconsin, Blood Research Institute, 8733 Watertown Plank Rd., Milwaukee, WI 53226, USA
Katherine Schulz
Affiliation:
Blood Center of Wisconsin, Blood Research Institute, 8733 Watertown Plank Rd., Milwaukee, WI 53226, USA
Richard W. Cole*
Affiliation:
Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY 12201, USA
*
Corresponding author. E-mail: [email protected]
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Abstract

The days of being able to ascertain instrument performance by simply peering through the eye pieces at a specimen are gone. However, users and granting agencies need to be confident that data collected on these instruments is uniform and quantifiable both over time and between instruments. Ideally, a LASER should not fluctuate, illumination should be completely uniform, and colors should be perfectly aligned. To check the current performance of imaging equipment, we conducted a worldwide research study utilizing three image-based tests: long-/short-term illumination stability, co-registration of signals across various wavelengths, and field illumination uniformity. To differentiate between “acceptable” and “unacceptable” performance, the deviation in illumination power could not exceed 10% (long term) or 3% (short term), the difference in the center-of-mass of imaged multicolored beads could not exceed >1 pixel between different wavelengths, and field illumination values could not exceed 10% (horizontal) or 20% (diagonal) deviation. This study established the current state of microscope performance through simple, efficient, and robust tests, while defining relative standards to assist cores in maintaining their instruments in optimal operating conditions. We developed cross-platform performance standards that will improve the validity of quantitative measurements made using various light microscopes.

Keywords

confocalstabilityperformance testingwide fieldco-registrationfield illumination
Type
Technology and Software Development Light and Confocal Microscopy
Information
Microscopy and Microanalysis , Volume 17 , Issue 4 , August 2011 , pp. 598 - 606
Copyright
Copyright © Microscopy Society of America 2011

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