| Light Microscopy Research Group (LMRG) |
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Membership History |
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| LMRG mission statement
Our goal is to promote scientific exchange between researchers, specifically those in core facilities in order to increase our general knowledge and experience. We seek to provide a forum for multi-site experiments exploring “standards” for the field of light microscopy. |
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| Current Membership |
Dr. Claire M Brown
(Chair) - McGill University
Ms. Carol J Bayles
- Cornell University
Prof. Richard Cole
- Wadsworth Center
Anne-Marie Girard
- Center for Genome Research and Biocomputing, Oregon State University
Dr Karen Martin
- West Virginia University
Dr. George McNamara
- U Miami
Cynthia Opansky
- BloodCenter of Wisconsin
Katherine Schulz
- Blood Center of Wisconcin
Marc Thibault
- Ecole Polytechnique
Dr Karen R Jonscher
(Ad hoc) (EB ) - University of Colorado Denver
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| Studies |
| 1) |
CURRENT STUDY: SPATIAL AND SPECTRAL RESOLUTION
Our second study was started one year ago and has two aims. The first is to test the resolution and quality of the laser scanning confocal microscope using point spread function measurements of sub-resolution fluorescent beads. The second is to test the accuracy of spectral detection calibration and spectral un-mixing in the confocal software.
Participants will receive samples of sub-microscopic fluorescent beads along with detailed protocols on how to measure the PSF and determine if the system is preforming ideally or needs alignment or if objective lenses are damaged. For the spectral studies a mirror slide will be provided and the reflection of the laser lines on the system into the spectral detector will be used to test accuracy. Finally, samples with beads labelled in the core with one orange dye and on the surface with a second orange dye will be used to determine the accuracy of the spectral un-mixing software.
Over 130 labs have signed up for the study and we will collect and assemble this data over the fall of 2011.
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| 2) |
RATIONAL FOR OUR STUDIES
Light microscopes have had a seminal influence on science for more than 300 years. The past three decades have seen a dramatic resurgence in the use of the light microscope, as well as very substantial technical advances in the field of light microscopy. This has in turn led to an increase in the use of the light microscope as a research tool. The most important advance has been the development of the confocal microscope, which combines the detection efficiency of fluorescence with the high resolution of the light microscope.
Improvements in design of optical components include, for example, aberration-corrected objectives (correction of both chromatic and spherical aberrations), more efficient filters (glass & AOBS), and improved detection (PMTs, CCD cameras and single photon avalanche diodes). As a result of these improvements, as well as improved performance and functionality of the systems, there has been a dramatic increase in costs of these types of instruments. The increase in cost coupled with decreasing grant support for research has resulted in many of these new instruments to be placed in multi-user facilities, i.e. imaging “cores”.
Establishment of imaging cores has led to a shift in responsibility for instrument acquisition, maintenance and training, from an individual PI to the director of the core and core personnel. Among the myriad of functions of the core is performance testing of the instrumentation. Users need to be confident that data collected will be uniform over time and between specimens. There is a need to developed standard Good Operating Practice (GOP) procedures for the imaging instrumentation. To this end, amongst other topics, the panel will discuss: (1) various aspects of “performance” test /GOP and an ongoing multisite study, (2) image format and who is responsible for storage, (3) oversight of data collection and image analysis; (4) keeping up with the newest technologies.
- proposed tests
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| 3) |
PAST STUDY ON LASER STABILITY AND CONFOCAL ALIGNMENT
We have data on the level of performance for various microscopes, mostly confocal. In our first study we provided laboratories across the world with plastic fluorescent slides for measuring short term and long term laser stabilities. These slides were also used to test for confocal alignment by looking at fluorescence image field uniformity. Finally, multi-colour bead samples were distributed to measure the co-registration of images collected in multiple confocal channels for different colour signals coming from the same bead. This study was very well accepted by the community and is now published:
"Quality assurance testing for modern optical imaging systems."
Stack RF, Bayles CJ, Girard AM, Martin K, Opansky C, Schulz K, Cole RW.Microsc Microanal. 2011 Aug;17(4):598-606. Epub 2011 Apr 11.
- The 39 Steps: A Cautionary Tale about “quantatativ
(28K)
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| 4) |
MIDL testing
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| Activities |
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| Protocols |
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| Publications |
| 1) |
Measuring and interpreting point spread functions to determine confocal microscope resolution and ensure quality control
- Nature Protocols Paper
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| 2) |
Quality Assurance Testing for Modern Optical Imaging Systems. Results of the LMRGs first study on laser stability, field uniformity, and co-registration of multi-colour confocal images.
- Microsc Microanal. 2011 Aug;17(4):598-606.
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| Membership History |
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