Multiphoton Microscope

Zeiss LSM 510 NLO Meta

Overview

A multiphoton (two-photon) laser scanning confocal microscope: Zeiss LSM 510 Meta NLO system with a Coherent Chameleon Ti:Sa Laser, coupled to an upright Zeiss Axioskop 2 microscope.  Components of the confocal include:

LSM 510 Meta NLO system:
– Two fluorescence channels (descanned)
– Two fluorescence channels (non-descanned)
– DIC transmitted channel
– Maximum scan resolution 2048 x 2048 pixels, 12 bits
META Accessory
– 32 channels divided by emission spectrum (377-719 nm)
Near Infrared femtosecond pulsed laser for multiphoton microscopy
– Coherent Chameleon system allows rapid computer-controlled tuning from 690-1040 nm
Conventional confocal laser lines:
– Ar laser 458, 488, 514 nm, 25 mW
– HeNe laser 543 nm, 1 mW
– HeNe laser 633 nm, 5 mW
AXIOSKOP 2 FS
– Fixed stage design for electrophysiology
– Motorized z-axis control
– Epifluorescence
– DIC
Zeiss LSM software
– Free download: Zeiss LSM Image Browser

Usage

1. The Zeiss LSM510 META NLO multiphoton imaging system is available to members of the Washington University Medical School and scientists from other research institutions, both educational and commercial, who have completed a basic training session and reserved a time slot.
2. Training sessions can be scheduled by contacting Kris (362-4876, hyrck@neuro.wustl.edu). The training session may be attended by 1-3 people and takes about two hours. The mandatory training covers the procedures of the Alafi Neuroimaging Laboratory, introduction to the principles of confocal/multiphoton microscopy and basic operations of the microscope controlled by Zeiss AIM software. Optional advanced sessions covering specialized aspects of imaging (spectral imaging, FRET, etc.) are available upon request. There is a flat fee of $100 for each training session. After completing the mandatory training session, the user is granted access to the facility and can work independently.
3. Users can reserve time slots on “first come first serve” basis using our web-based calendar (www.calendar.yahoo.com; Login ID: alafi2photon, contact Gary or Kris for password). When signing up, please put down just your computer login ID into the timeslot’s “TITLE” field. Other information (for your own use) may be put into the calendar’s “NOTES” field for that time slot.
4. The users might want to check the calendar for urgent messages regarding the instrument status and staff availability.
5. Assistance in solving imaging problems and answering application questions is readily available by contacting Kris (lab: 362-4876, hyrck@neuro.wustl.edu) or Gary (cell: 314-647-1812, microscope@zootalk.net) during normal business hours (9 a.m.-5p.m.).
6. Users planning to work after hours or on weekends should make sure that they have everything they need (room access, working passwords, etc.) to complete their tasks prior to scheduling their work as staff assistance might not be available.
7. All comments and concerns regarding our facility, equipment and procedures should be directed to Kris or Gary.
8. To allow a fair and efficient utilization of the microscopes, we recommend that:
   1. The microscope should be reserved in advance for the time that is needed to perform the desired tasks.
   2. Time reservation gives the user absolute priority over unscheduled/extended usage. Still, showing some flexibility would be greatly appreciated.
   3. Time reservations that cannot be utilized should be cancelled as soon as possible to let other users use all available time. Reservations not cancelled by 12:00 pm (noon) on the preceding day will be subject to “reserved/unused” time charge corresponding to 50% of normal rates.

More information

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