CORE B:
IMAGE ANALYSIS CORE

Director: Asma Nusrat

Co-Director: Ifor Williams

Equipment available:

1. Confocal Microscopes (2), Zeiss 510 with META system equipped with Argon and HENE/3PMT lasers. One confocal microscope housed in the Neuroscience department is equipped with a Ti-sapphire laser for two-photon excitation microscopy and live tissue imaging in addition to UV lasers. Further refinements in the software program are planned over the next five years.
2. Flow Cytometer, Becton Dickinson FACSCalibur flow cytometer capable of four-color analysis.

     3.   Additional facilities

• Nikon upright fluorescence microscope
• Cryostat for generating frozen sections
• Additional complete work station with Zeiss computer software and Metamorph program for image analysis.
• Automatic tissue processor for paraffin embedment and tissue sectioning.                         
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Image Analysis Core

The Image Analysis Core encompasses two state-of-the-art Zeiss LSM 510 confocal microscopes equipped with META detectors and Argon and HENE/3PMT lasers. One confocal microscope housed in the neuroscience department is additionally equipped with a Ti-sapphire laser for two-photon excitation microscopy and live tissue imaging in addition to UV lasers.

The confocal microscopes are presently fully functional and funding from this grant will subsidize the user cost of this core facility. Both microscopes are presently being utilized by core members. In addition to the confocal microscope, the Image Analysis Core will provide a facility to generate frozen sections. A technician will oversee and manage the confocal microscope and core-related issues. This includes training of new users, troubleshooting, maintaining the schedule and payment structure set up for use of the confocal microscope. Scheduling of confocal microscope use will be done on a web-based calendar. The core has a billing system based on hourly use of the confocal microscope.

User Structure:

1. Center members (29 members)
2. Faculty in the Department of Pathology
3. Outside users

Time Allocation:

95% of total time will be allocated to Core members and Pathology Department Users. Outside users will have limited access to this facility for 5% of its operational time.

Confocal Core Charges:

Cost:  $35/hour

This amount will be subsidized for core users to $22/hour

Additional Facilities:

The use of a cryostat, additional microscope, work station and the automatic tissue processor will not be billed. This equipment will be maintained as a part of the core facility.

Flow Cytometry Core

The Flow Cytometry Core consists of a Becton Dickinson FACSCalibur flow cytometer (2 lasers; 4 color capability) with integrated FACSLoad unit and a separate G4 PowerMac computer workstation running CellQuest and FlowJo for offline flow data analysis.  Initial training of new users is done with the assistance of personnel of the Medical School Flow Cytometry Core.  Assistance with problems arising during experiments is provided by a technician who also performs recommended routine maintenance on the instrument and arranges for service calls covered by the service contract when these are needed.  Scheduling of usage of the flow cytometer is done by a web-based system.  Billing is based on the time of usage which is tracked by the computer that controls data acquisition by the flow cytometer.

User Structure:

1.  Center members (29 members) -- currently about 35% of usage

2.  Faculty in the Department of Pathology -- currently about 30% of usage

3.  Outside users -- currently about 35% of usage

Time Allocation:

At this time, Center members and faculty from the Department of Pathology have not experienced any difficulties in scheduling the instrument which is available to many users 24/7.  If booking of the instrument by outside users increases to the point where user groups #1 and #2 are unable to book adequate amount of time on the instrument, restrictions on the time usage by outside users will be instituted.

Flow Cytometry Core Charges:

Cost: $21/hour for DDRDC members.