The LSM 410 confocal microscope is located on the sixth floor of the Medical School in room E638. The system consists of a Zeiss LSM 410 laser-scanning module mounted on a Zeiss Axiovert 100 microscope. The system offers five excitation wavelengths (633nm, 568nm, 488nm, 364nm, and 351nm), and for detection, three separate PMTs, each with its own adjustable pinhole and emission filter wheel. A Bioptechs Delta T4 heating stage and objective heater are available to facilitate live cell imaging on all the Zeiss systems. For additional information and training, please contact Maryanne Pendergast at x2575.

• Maryanne Pendergast
  Training and Facility Manager
  368-2575 (office)
  216-224-8649 (cell)
  mxp19@case.edu

• Zeiss LSM 410 laser scanning module
  
• Zeiss Axiovert 100 inverted microscope
  
• Internal HeNe laser, ex 633nm
  
• External Omnichrome Ar/Kr laser, ex 488nm/568nm respectively
  
• External Coherent Enterprise UV laser, ex 351nm and 364nm
  
• 3 PMTs for simultaneous detection of three separate channels
  
• Wide assortment of high quality objectives with both short and long working distances
  
• Bioptechs Delta T4 temperature control system for live cell imaging
  

• All prospective users are expected to have experience with basic epi-fluorescence microscopy including standard fluorescent or GFP techniques.
  
• Everyone must be trained and approved for independent operation of the system by Maryanne. The average training time is approximately 5-6 hrs, and is related to the user’s experience with microscopes and PC computers.
  
• Everyone is expected to use and care for the equipment as demonstrated during the initial training session. Abuse of the system may result in the cancellation of user privileges; repair costs will be charged to the lab at fault.
  
• Confocal assistance is available at an hourly rate for one-time evaluation experiments and infrequent users.
  
• Data can be stored short-term on the NIC server which is accessible to all computers within the facility as a mapped drive (Z:\); the server is available to other computers through a web interface. Data can also be saved to CDs and 100MB ZIP drives.
• Sign up during the day is first come/ first serve and may be restricted to two half days per week during exceptionally busy times.
  
• Additional time is available after hours and on the weekend for experienced users (reduced rate, first come/first serve).
  
• Sign up should include user and PI names, time, and phone number.
  
• After your session please check the sign-up calendar:
  
  1. If the next user is scheduled within three hours, please leave the laser and Hg arc lamp on and turn the power adjust knob to minimum (6 o’clock).
  2. If the next user is scheduled to arrive after three hours, please turn the system off.
  3. If you are the last user of the day, and someone is scheduled to use the system after hours, please call to ascertain that he or she will actually be coming. This is especially important for Friday evenings.

• Want to know how a microscope works? About color and light? Here is a site that has it all, including interactive java tutorials that are both educational and FUN!
  http://micro.magnet.fsu.edu/primer/index.html

• Concerned about possible spectral overlap between fluorescent dyes and resultant bleed-through problems? This interactive site will allow you to superimpose the excitation and emission spectra of multiple fluorochromes on a normalized axis. Use this site to plan the fluorescent dyes for your experiments.
  http://www.microscopy.bio-rad.com/fluorescence/fluorophoreDatab.htm
  

• The confocal listserv is maintained by the University of Buffalo and has archived discussions dating from November 1991. Topics covered include anything and everything having to do with confocal microscopy. It’s great for trouble shooting problems, and also for the breadth of information available. A visit to this site is a must for all confocal users!
  http://listserv.acsu.buffalo.edu/archives/confocal.html

• Website for Carl Zeiss, North America
  http://www.zeiss.com/micro

Zeiss LSM 410 Quick Start Guide

CWRU Laser Safety Manual

OSHA Laser Safety