Training: All users on the PINC Leica Confocal Microscope are required to have personally completed the introductory training and hands on training sessions from Dr. Neal Kramarcy and to have interviewed with Dr. Robert Sealock before independently using the LeicaTCS-NT confocal system.  Training is conducted on Mondays by appointment.   The purpose of training is to develop independent, knowledgeable confocal users.

• Introductory Training: Tutorial (~4 hours).  Maximum of 4 users at one time.  A test sample will be scanned under the confocal.

  • Quick tour of the confocal system
  • Laser and electrical safety
  • "Only users trained by us may drive system" must see Dr. Robert Sealock before using system solo
  • Safe and knowledgeable operation of Leica DM IRBE microscope
    • Ensure microscope is off before powering up mercury lamp
    • Before changing slide ensure that focus is at lowest position
    • Only use the objective changer control when focal plane is found and upper limit is set at that focal plane.
    • Objective may only be changed manually when focus is at lowest position
    • Krypton & Argon Laser power knobs at minimum, then key on laser

      Leave laser at minimum power if next user is arriving within next three hours

    • Never cycle lasers on and off
  • Powering up the confocal system
  • Logging onto WindowsNT and running LeicaTCS-NT
  • Principal of confocal microscopy
  • Scanning - objective, pinhole, laser power, detector sensitivity
  • Saving images - a brief word about multi-page TIFF image files
  • Powering down and next user consideration
  • Cleanup

• Hands on Training: (~2 hours).  Scan with user's sample.  Preferably with individual users or groups wishing to work on same type of fluorescence.

  • Powering up the system
  • Operation of Leica DM IRBE
  • Scanning multiple channels
  • Signal level and power - PMT gain - pinhole - noise
  • z-series and/or x-z scanning
  • Etc. as fits user's experiment, e.g. time lapse, DIC & transmitted light
  • File storage - multichannel tiff image issues - cleanup files after 1 week.
  • Network transfer of data
  • Powering down
  • Cleanup

• 3^rd Training: (optional) Image processing or advanced scanning

  • Multichannel tiffs, splitting multichannel tiff files & PhotoShop etc.
  • Sequential scanning
  • Image enhancement
  • Z-series handling
  • Rendering
  • Extended focus & 3D-reconstructions
  • User requests

• User to interview with Dr. Robert Sealock - a word about user responsibilities

• User issued with WindowsNT password and web booking page booking ID number, and instruction of the use of the world wide web booking system with Dr. Neal Kramarcy.

• User/Lab. may be issued room key (deposit $20).

  Questions? Contact  Neal at kramarcy@med.unc.edu or room 158 M.S.R.B. (2 floors above confocal cave room).

  Last updated 31/07/03

					© 1998-2003 Michael Chua, Cell & Molecular Physiology, UNC.  All rights reserved.
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