LOCATION:
Rooms LC-207, E-135
STAFF:
Frederick R. Maxfield, Ph.D., Professor & Chairman, 212-746-6405
Lynda Pierini, Ph.D., Director, 212-746-6459
Leona Cohen-Gould, Manager, 212-746-6146; page: 746-6700 #17334
FACILITY
DESCRIPTION:
The Optical Microscopy Facility provides equipment and expertise
to assist investigators in applying sophisticated light microscopy
techniques to their research. The equipment in this facility includes
a confocal microscope to be used for 3D image acquisition, and
Dell and Silicon Graphics workstations to quantify, manipulate
and render 3D fluorescence microscopy images. In addition to access
to the equipment, the Facility provides consultative and collaborative
assistance in research efforts. This allows researchers with expertise
in biochemistry and molecular biology but who are unfamiliar with
optical microscopy methods to apply these techniques in their
research. The facility Manager and Director provide advice in
the design of experimental programs, instruction in the use of
the equipment, and assistance in the interpretation of data.
EQUIPMENT:
The
facility now has a Zeiss LSM510 laser scanning confocal microscope.
This microscope has four detectors (3 for fluorescence 1 for bright
field/Differential Interference Contrast). It is illuminated by
3 lasers (Argon, "green" Helium-Neon and "red" Helium-Neon)which
supply excitation lines at 458nm, 488nm, 514nm, 543nm, and 633nm,
allowing simultaneous confocal imaging of 3 fluorophores (e.g.:
FITC, Rhodamine and CY5) plus the bright field/DIC image. The
facility also has a Dell workstation that runs the Zeiss proprietary
image analysis and 3-D rendering software as well as a Silicon
Graphics O2 workstation running Image Space 3-D image analysis
software (Molecular Dynamics) and software for protein structure
analysis.
ORGANIZATION
AND SERVICES PROVIDED
1.
CONSULTATION. Consultation on experimental design and
approach is provided by the Facility Manager and Director.
2.
CONFOCAL MICROSCOPY IMAGING. The investigator is expected
to have thoroughly discussed their experimental plan with the
facility manager or director before preparing samples for imaging.
This is to ensure feasibility of the project, and that the investigator's
sample preparation can be imaged by the confocal microscope.
Users will be responsible for their own sample preparation.
The
facility can acquire images of fluorescently labeled tissue
or cells where the background fluorescence from out of focus
objects has been reduced. If needed, optical sections can be
obtained to image the entire cell volume. A wide variety of
fluorophores that emit in the green and red portions of the
spectrum can be imaged, such as: FITC, GFP, Alexxa 488, Cy3,
Rhodamine, Texas Red, DiI, Cy5 (to name a few). Once acquired,
these images can be quantified, or manipulated in 3-D on the
facility's Dell and Silicon Graphics workstations.
Investigators
will be responsible for archiving their data.
Investigators
will be allowed to use the facility unassisted only after they
have been trained to the Manager's satisfaction. Otherwise,
the manager will assist in the facility's usage. Misuse of the
instrument will result in charges for repairs and revocation
of permission to use it.
3.
TRAINING. The Manager will provide training in usage
of the confocal microscope and the workstations. Training sessions
are arranged in consultation with the Facility Manager.
PART II: FEES FOR SERVICE
The
fees for the services of the Optical Microscopy Facility were
set as follows on January 1, 2000.
All
users must provide an account to be charged before using the facility.
Signed up time is charged whether or not it is used unless a replacement
user is found.
Each
research group will receive up to 6 hours of initial training
without added charges. Additional training will be available at
the assisted use rate.
Confocal
microscopy charges
| Independent use: |
$50 per hour Monday - Friday 9 am to 6 pm |
|
$25 per hour Monday - Friday 6 pm to 9 am |
| Assisted use: |
$75 per hour |
Image
processing and display computers
| Independent Use: |
$10 per hour (max. $20/day); overnight use: $20 |
| Assisted use: |
$40 per hour |
All
users are responsible for properly maintaining the microscope.
Damage or problems must be reported promptly to the facility staff.
Research groups may be charged for repairs caused by improper
use including failure to clean optics properly. Pre-existing problems
must be reported immediately to avoid being charged. Experience
indicates that these repairs can cost as much as $4000 for careless
treatment of objectives.
PART III: PROCEDURES FOR USE OF THE OPTICAL MICROSCOPY FACILITY
PERSON
TO CONTACT:
Leona Cohen-Gould, 746-6146
746-6700 #17334 (pager)
SCHEDULING:
The facility is open Monday-Friday 9 am-5 PM, and after hours
at the discretion of the staff. Scheduling is done by sign-up
sheet and coordinated with Ms. Cohen-Gould for those needing
assistance. Users will be billed for reserved time unless it
is canceled at least one day in advance or used by another group.
BILLING:
Billing will be generated at the end of each month and processed
through the Biochemistry Department.
ZEISS LSM 510 CONFOCAL SIGN-UP RULES
IF
THE LASERS HAVE BEEN SHUT OFF, THEY MUST COOL FOR 2 (TWO) HOURS.
Check the schedule to see if someone is booked after you. If
yes, leave the microscope running.
