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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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References

REFERENCES

Bustin, S.A., Benes, V., Garson, J.A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M.W., Shipley, G.L., Vandesompele, J. & Wittwer, C.T. (2009). The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55(4), 611–622.CrossRefGoogle ScholarPubMed
DeRose, P.C. & Resch-Genger, U. (2010). Recommendations for fluorescence instrument qualification: The new ASTM Standard Guide. Anal Chem 82(5), 2129–2133.CrossRefGoogle ScholarPubMed
Guterman, L. (2010). Access sparks action. NCRR Reporter 34(1), 4–8.Google Scholar
Latterich, M. (2006). Publishing proteomic data. Proteome Sci 4, 8.CrossRefGoogle ScholarPubMed
Nesvizhskii, A.I., Vitek, O. & Aebersold, R. (2007). Analysis and validation of proteomic data generated by tandem mass spectrometry. Nat Methods 4(10), 787–797.CrossRefGoogle ScholarPubMed
Pawley, J. (2000). The 39 steps: A cautionary tale of quantitative 3-D fluorescence microscopy. BioTechniques 28(5), 884–886.CrossRefGoogle ScholarPubMed
Pawley, J. (2006). Handbook of Biological Confocal Microscopy. New York: Plenum.CrossRefGoogle Scholar
Reiss, S.M. (2010). Quality and standards: Making bioimaging measure up. BioOptics World 3(1), 14–18.Google Scholar
Rosenthal, C.K. (2009). Light microscopy: Contrast by interference. Nature Milestones | Milestone 8.Google Scholar
Sharma, D. (2010). Standardized units for reproducible imaging experiments. Biophotonics (September), 32–35.Google Scholar
Zucker, R.M. (2006a). Quality assessment of confocal microscopy slide-based systems: Instability. Cytometry A 69(7), 677–690.CrossRefGoogle ScholarPubMed
Zucker, R.M. (2006b). Quality assessment of confocal microscopy slide based systems: Performance. Cytometry A 69(7), 659–676.CrossRefGoogle ScholarPubMed
Zucker, R.M. & Price, O. (2001). Evaluation of confocal microscopy system performance. Cytometry 44(4), 273–294.3.0.CO;2-N>CrossRefGoogle ScholarPubMed