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Light Microscopy Research Group (LMRG)

Mission

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.

Current Members

Benjamin Abrams - University of California Santa Cruz

Constadina Arvanitis - Northwestern University

Linda Callahan - University of Rochester Medical Center

Lisa Cameron - Duke University

Richard Cole - Wadsworth Center

Joseph Dragavon - University of Colorado

David Kirchenbuechler - Northwestern University

Kristopher Kubow (Chair)James Madison University

Gary S Laevsky - Princeton University

Valeria Mezzano - New York University School of Medicine

Thomas Pengo - University of Minnesota

James A Powers - Indiana University 

Joshua Zachary Rappoport - Northwestern University

Mark Sanders - University of Minnesota 

Nelly Vuillemin - McGill University

Erika Wee - McGill University

Nancy Fisher (EB Liaison) - University of North Carolina at Chapel Hill School of Medicine

Studies

1)The third study of the Light Microscopy Research Group (LMRG) is aimed at creating a 3D biologically relevant test slide and imaging protocol to test for: (1) System resolution and distortions in 2D and 3D. (2) The dependence of Intensity quantification and image signal-to-noise of the microscope on imaging depth. (3) The dependence of the microscope sensitivity on imaging depth. The test sample consists of a mixture of fluorescence microspheres imbedded in a 120 um-thick layer of CyGel (BioStatus, UK, Cat# Cy10500) with a refractive index of 1.37 closely matched to biological tissue. A double-sided adhesive 18 mm square spacers with a well (9 mm diameter, 120 um deep) were used for the sample preparation (Electron Microscopy Sciences, Cat# 70327-8s). The mixture of microspheres includes 1 um Orange, 2.5 um Green 20% brightness, 2.5 um Green 100% brightness, 6 um Far Red 36%, 6 um Far Red, 15 um Blue Core/Orange Ring.

Activities

1)ABRF2014 LMRG Group and CCMA Travel Award Winners
    - Richard Cole, John Russ, and Claire Brown (2,464K)
    - CCMA Travel Award Winners (1,934K)
2)ABRF2013 LMRG Photo
    - ABRF2013 LMRG Photo 
3)ABRF 12 RG group photo
    - 2012 Meeting Group Photo 
4)ABRF 12 RG intro talk
    - LMRG 2012 Overview Talk 
5)ABRF 12 RG data talk
    - Study #2 Presentation (7,190K)
6)Proteomics workshop-- Get on Your Way to Microproteomics with laser Microdissection
    - Proteomics - Sarah Baxter 
7)Proteomics workshop-- Sample prep
    - Tissue Proteomics Sample Preparation Leica Talk 
8)ABRF 11 LMRG group photo
    - View Document (3,315K)
9)ABRF 11 RG talk
    - View Document (13,284K)
10)ABRF 2010 talk
    - View Document (4,643K)
11)Talk for ABRF 09
    - View Document (10,567K)

Protocols

1)This is a paper written by Cole and Brown and gives a lot of background on how to measure point spread functions, why you may want to measure them, how to prepare bead slides and then a detailed protocol for setting up the instrument and measuring and interpreting the PSF. This articile should be looked at first before referring to the specific protocols for the individual microscope platforms.
    - Nature Protocols Paper (1,371K)
2)PSF Protocol Leica SP5 - Measuring the PSF with the 0.175 um green fluorescent bead sample.
    - PSF Protocol Leica SP5 
3)PSF Protocol Nikon A1 - Measuring the PSF with the 0.175 um green fluorescent bead sample.
    - PSF Protocol Nikon A1 
4)PSF Protocol Olympus FV1000 - Measuring the PSF with the 0.175 um green fluorescent bead sample.
    - PSF Protocol Olympus FV1000 
5)PSF Protocol Zeiss 510 - Measuring the PSF with the 0.175 um green fluorescent bead sample using the AIM software.
    - PSF Protocol Zeiss 510 
6)Spectral Accuracy Protocol Leica SP5 - A mirror slide will be provided and you will look at the reflections of the laser lines into the spectral detector to determine if the wavelength readouts are accurate.
    - Spectral Accuracy Protocol Leica SP5 
7)Spectral Accuracy Protocol Olympus FV1000 - A mirror slide will be provided and you will look at the reflections of the laser lines into the spectral detector to determine if the wavelength readouts are accurate.
    - Spectral Accuracy Protocol Olympus FV1000 
8)Spectral Accuracy Protocol Zeiss 710 - A mirror slide will be provided and you will look at the reflections of the laser lines into the spectral detector to determine if the wavelength readouts are accurate. Uses the ZEN software.
    - Spectral Accuracy Protocol Zeiss 710 (261K)
9)Spectral Accuracy Protocol Zeiss 510 - A mirror slide will be provided and you will look at the reflections of the laser lines into the spectral detector to determine if the wavelength readouts are accurate. Uses the AIM software.
    - Spectral Accuracy Protocol Zeiss 510 
10)Spectral Separation Accuracy Protocol Leica SP5 - Protocol for spectral separation testing of software un-mixing with double orange stained beads.
    - Spectral Separation Accuracy Protocol Leica SP5 
    - Bead Core Spectra Text File 
    - Bead Ring Spectra Text File 
    - Bead Core Spectra lsf File (12K)
    - Bead Ring Spectra lsf File (9K)
11)Protocol for testing the spectral separation accuracy of the Olympus FV1000.
    - Spectral Separation Protocol Olympus FV1000 (1,133K)
12)Study #1 Laser Stability, Alignment and Co-registration Protocol
    - Study #1 Laser Stability, Alignment and Co-registr 
13)Improved GFP imaging in live cells
    - Improved GFP imaging in live cells 

Membership History

Member NameOrganizationDetails
Pamela Scott AdamsTrudeau Institute Ad hocEB Liaison: 02/09 - 03/10
Ms. Carol J BaylesCornell University Member: 04/08 - 05/15
Dr. Claire M BrownMcGill University Chair: 03/11 - 04/15
Member: 09/10 - 03/18
Prof. Richard ColeWadsworth Center Chair: 03/08 - 03/11
Dr. Arnold M. FalickHHMI-UC Berkeley Ad hocEB Liaison: 03/08 - 02/09
Dr. Paul FurcinittiUMass Medical School Member: 05/13 - 12/14
Anne-Marie GirardCenter for Genome Research and Biocomputing, Oregon State University Member: 02/10 - 05/15
Dr William G HendricksonUniv. of Illinois, Chicago EB Liaison: 09/13 - 04/14
Dr Karen R JonscherUniversity of Colorado Denver Ad hocEB: 04/10 - 03/13
Dr Karen MartinWest Virginia University Member: 12/08 - 05/15
Dr. George McNamaraU Miami Member: 06/11 - 03/13
Ad hoc: 03/13 - 12/14
Ms. Kary OakleafMolecular Probes, Life Technologies Member: 06/13 - 12/14
Cynthia OpanskyBloodCenter of Wisconsin Member: 02/09 - 12/13
Katherine SchulzBlood Center of Wisconcin Member: 03/09 - 04/13
Robert F. StackWadsworth Center NYSDOH Member: 02/09 - 03/11
Marc ThibaultEcole Polytechnique Member: 06/11 - 03/14
Dr. Erika WeeMcGill University 

Member: 05/13 - 04/18

Chair:04/15-04-18

Dr. Frances Weis-GarciaMemorial Sloan Kettering Cancer Center EB Liaison: 05/14 - 04/15

 

Publications

  1. Microsc and Microanal. 2013 Dec;19(6):1653-68.
    Cole, RW, Thibault, M, Bayles, CJ, Eason, B, Girard, AM, Jinadasa, T, Opansky, C, Schulz, K, Brown, CM,
  2. Microsc Microanal. 2011 Aug;17(4):598-606.
    Stack RF, Bayles CJ, Girard AM, Martin K, Opansky C, Schulz K, Cole RW.Microsc Microanal.
  3. Nature Protocols 6 (2011):1929
    RW Cole, T Jinadasa, CM Brown
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