Microscopy and Imaging Shared Resource

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Olympus Fluoview-FV300 Laser Scanning Confocal System

Olympus IX-70Olympus IX-70 microscope


Upgraded in January 2005 with inverted Olympus IX-70 microscope and six laser lines (blue diode/405nm, Argon/458, 488, 514nm, HeNe/543nm, HeNe/633nm, all with AOTF for fast individual intensity control) for triple color imaging or live imaging in two colors. New capabilities include FRET for CFP/YFP and CFP photo activation in living cells.

Software:

  • Fluoview acquisition software including Timelapse and Ratiometric modules

Objectives:

  • Long working distance (samples in dishes, use phase contrast for bright field imaging) or high resolution short working distance condensers (coverslips on slides or coverslips on the bottom of multi-well slides-Nomarski/DIC for bright field imaging)

  • Dry lenses with long working distances (10X, 25X, 40X)

  • Oil lenses (40X N.A. 1.0, 60X N.A. 1.4, 100X N.A. 1.3)

  • 60X NA 1.2 Water lens for live imaging

Other Capabilities:

  • Bioptechs regulated temperature Delta T open dish system and objective heater or Bioptechs (FCS2) regulated temperature closed chamber system with perfusion pump and objective heater.

  • Transmitted light detector for Nomarski/DIC images

  • Live imaging, time lapse imaging of live cells in glass bottom dishes (Bioptechs Delta T or FCS2 systems) or plastic multi-well plates, in fluorescent or transmitted light

Multi-color Fluorescence Imaging with Quadruple Band Pass:

  • DAPI/FITC/Texas Red/Far Red

Single Band Pass Filters:

  • FITC: (excitation=450nm, dichroic=505nm, emission=535nm)

  • Texas Red: (excitation=560nm, dichroic=595nm, emission=645nm)

  • CY5: (excitation=620nm, dichroic=660nm, emission=700nm)

  • DAPI: (excitation=360nm/405 nm -- Hg Lamp/Blue diode Laser, dichroic=400nm, emission=460nm)

Specifications for Dichroics and Filters for Laser Confocal Imaging

  • Download a chart of Filters/DM configurations

Computer & Imaging Software:

  • Pentium IV running Windows 2000, 1GB RAM, 70GB Hard Drive, writable DVD/CD-ROM

  • Olympus Fluoview FV300 version 3C Acqusition Software

Acknowledgments:

PLEASE REMEMBER: For funding purposes, it is important to acknowledge the Microscopy & Imaging Shared Resource Facility in all publications that include data derived from the Facility. For Olympus Fluoview Laser Confocal microscope, this should include the following statement:

"This work was supported in part by the Lombardi Comprehensive Cancer Center Microscopy and Imaging Shared Resource funded by U.S. Public Health Service Grant 2P30-CA-51008, 1S10 RR15768-01, 1 S10 RR019291-01A2, and the Carey Lackman Slease Fund."

If you have comments or suggestions, email Michael Johnson, director of MISR at Johnsom@georgetown.edu.