The Leica TCS SP2 MP confocal microscope is a state of the art system that combines spectrophotometric detection with confocal microscopy. Instead of traditional dichroic filter sets, a prism is used to separate the emitted fluorescence, resulting in a very flexible and efficient detection system The scan head is mounted on a Leica DMIRE2 inverted microscope. For excitation the system offers six visible wavelengths (458nm, 476nm, 488nm, 514nm, 543nm, and 633nm) plus a tunable (720nm-950nm) IR light source (Choherent Chameleon XR MP laser). The system is located in the Neuroscience department, room E639. Please contact Yoshi Imanishi for training and additional information about this system.

• Dr. Yoshikazu Imanishi
  Assistant Professor, Pharmacology
  368-5226
  yxi19@case.edu

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

• Leica TCS SP2 laser scanning system
• Leica DMIRE2 inverted microscope
• Multi-line Argon laser (458nm, 476nm , 488nm, 514nm)
• HeNe laser (543nm)
• HeNe laser (633nm)
• Multi-photon Coherent Chameleon XR laser, tunable (720-950nm)
• Stage and objective heater available by special arrangement

• All prospective users are expected to have experience with basic epi-fluorescence microscopy including standard fluorescent or GFP techniques.
  
• All new users need to go through basic microscopy training with Maryanne and should read the CWRU Laser Safety Manual.
  
• Everyone must be trained and approved for independent operation of the system by Yoshi and Maryanne. The average training time is approximately 3 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 not covered by the service contract 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, DVDs, and USB 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 one hour, please leave the HBO lamp on and turn the main argon laser to stand-by. The other lasers may be turned off.
  2. If the next user is scheduled to arrive after one hour, 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

Leica TCS SP2 User Manual

CWRU Laser Safety Manual

OSHA Laser Safety