The Huck Institutes of the Life Sciences

Olympus Fluoview 1000

A four-laser confocal microscope using auto-setup to install objectives, lasers, and detectors for unique dye combinations; FRAP (with SIM scanning), FRET, and spectral imaging are available.

Specifications

  • 4 lasers (405, 488, 543, 633 nm) plus 405nm SIMS for FRAP
  • Time controller for complex experiments

Details

Microscope:
Olympus IX70 inverted microscope with fluorescence burner
Objectives:
UplanAPO 10X/0.40, UPlanFL 20X/0.50, UAPO/340 40x/1.50 water, UPlanFL 40X/1.3 oil, PlanApo 60X/1.4oil, UPlanSApo 100X/1.4 oil
Lasers:
Four single-line lasers with individual shutters that are software-controlled for sequential acquisition.
  • Violet (405nm, 10mW)
  • Blue argon (488nm, 10mW)
  • Green HeNe (543nm, 10mW)
  • Red HeNe (633nm, 10mW)
Scanning modes:
  • 1-dimensional (spot-scan), 2-dimensional (X-Y vector, X-Z, or time)
  • 3-dimensional (X-Y-Z, or X-Y-time) or 4 dimensional (X-Y-Z-time)
Transmitted light:
  • External halogen light source connected to microscope via fiber cables, external PMT for light collection.
  • DIC optics are available for all objectives.
Resolution:
Confocal LSM improves the resolution of images by recording fluorescence or reflected light when a laser is focused at set focal planes in tissues and cells, rejecting all other light coming from planes above or below the one of interest. This improves resolution of sub-micron structures within fixed and live cells and tissues. A series of images can be recorded axially and either analyzed separately or incorporated into a composite image.
SIM Scanning:
  • Simultaneous photobleaching and scanning.
  • Excellent for rapid FRAP or acceptor pFRET.
Spectral imaging:
Using either established fluorescence spectra or blind spectral deconvolution Time controller: enables multiple diverse imaging conditions to run with automatic controls
Software:
Fluoview: captures files as MultiTIFF files (*.oib) and displays as TIFF files (*.tiff)
Output:
  • Files will be transferred to an off-line computer.
  • Users will transfer data via FTP or download it to a writeable CD-ROM.
  • Display TIFF images can be printed directly using graphics software (such as Photoshop or PowerPoint).
  • MultiTIFF images can be reanalyzed using the Fluoview software or ImageJ.
 
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Fluoview