Michael Hooker Microscopy Facility (MHMF.ORG)

User Training

Training: All users of the Michael Hooker Microscopy Facility are required to have personally completed introductory and hands on training from the facility staff for each piece of equipment.  Generally Training is conducted by appointment to small groups or individuals.   The goal of training is to develop independent, knowledgeable microscope users.  Hence, training is best undertaken close to the time when samples are soon to be ready and equipment first needs to be used.

• Introductory Training: Tutorial (~3 hours).  Ideally 2 and a maximum of 3 users at one time.  A test sample will be scanned under the confocal.

  • Each new user should fill out user information form on or before 1st training day   (http://mhmicroscopy.med.unc.edu/Users-new/default.html)
  • Optionally fill out billing information and logon account preferences & booking ID choice http://mhmicroscopy.med.unc.edu/Users-new/default-billing.html
  • Quick tour of the confocal system
  • Laser and electrical safety
  • "Only users trained by us may drive system"
  • Safe and knowledgeable operation of microscope
    • Ensure microscope is off before powering up mercury lamp
    • Ensure mercury lamp is cool before turning it on
    • 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
    • Argon Laser power knobs at minimum, then key on laser
    • Leave laser at minimum power or standby if next user is arriving within next 1 hour
    • Never cycle lasers on and off
    • Ask facility personnel if objective(s) needs to be changed
  • Powering up the confocal system
  • Logging onto Windows and running acquisition software
  • Principal of confocal microscopy - the emission pinhole
  • Scanning - objective, pinhole, laser power, detector sensitivity
  • Saving images
  • Cross talk/bleed through
  • Powering down and next user consideration
  • Cleanup

• Hands on Training: (~3 hours).  Scan with user's sample.  Preferably with individual users or groups wishing to work on same type of sample.

  • User issued with facility domain password and web booking page booking ID number
  • Instruction of the use of the world wide web booking system.
  • Powering up the system
  • Operation of microscope
  • Scanning multiple channels
  • Signal level and power - PMT gain - pinhole - noise
  • Bleed through (again)
  • z-series and/or x-z scanning
  • Etc. as fits user's experiment, e.g. time lapse, DIC & transmitted light
  • File storage - cleanup files after 1 month
  • Network transfer of data
  • Powering down
  • Free offline viewing software
  • Cleanup

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

  • User requests
  • Multichannel scanning file formats & PhotoShop etc.
  • Sequential scanning
  • Image enhancement
  • Z-series handling
  • Extended focus & maximum projection
  • 3D-viewing
  • Rendering
  • XZ scanning
  • Time lapse
  • FRAP
  • FRET

• After hours access using UNC SOM building card.  Card swipe access application go to http://microscopy.unc.edu web page

  Questions?  Contact facility personnel in 6007, 6033a or 6129 Thurston Bowles

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		Copyright 2001-2014  Dr. M. Chua, Program in Molecular Biology & Biotechnology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599
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