CELLS AND TISSUES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Cytometry and Cell Sorting Lab Udai P Singh, Leader Advanced cell sorting and flow cytometry analysis have broad multidisciplinary uses to conduct experiments in several areas of research including cell and developmental biology, immunology, HIV/AIDS, hematology, oncology and cardiovascular disease. Faculty oversight and dedicated full time technical support staff is facilitating access and ensure effective use of the Core's capabilities. The flow cytometry core is conceptualized as a set of enabling technologies available for program/project development. The flow cytometry is providing advanced and reliable technical support to MSM researchers as well as others institution using these cutting-edge technology platforms. The core is also significantly enhancing the research and training environment for undergraduate, graduate and medical students engaged in translational and clinical research at MSM. Service Charges: 1. Flow Cytometry on the FACScan $ 30.00/hour 2. Cell sorting on the FACStar $ 60.00/hour 3. Data analysis for MSM faculty Free 4. Data analysis other Institution Free Dr Singh started employment in this facility in September 2006, and in the first four months he has analyzed 450 samples from the various investigators. He has conducted 12 separate experiments by different investigators and run the FACScan approximately 24 hour in total. Dr. Singh is expecting more users in the next six month. Molecular Histology Lab John Patrickson, Leader This is a new core unit that will provide histology services to the MSM research faculty, graduate students, and the Atlanta University Center. This facility is equipped to provide basic histological services including: paraffin embedding, paraffin sections, frozen sections, immunohistochemical and H&E staining, and in situ hybridization. The facility consists of a paraffin tissue embedding center, rotary microtomes, cryostat, tissue floatation bath, microwave, tissue processor, and slide warmers. Service Charges The investigator is responsible for the fixation and blocking of the tissue. For routine fixation it is recommended that the tissue is fixed with a fresh solution of 10% neutral buffered formalin or paraformaldehyde. For best results, it is recommended that the fixatives be purchased in small quantities. (Contact the laboratory for details) For frozen tissue preparations contact the laboratory. Summary of Charges: Paraffin: Embedding $4.00; Cut, Stain, Coverslip -1st slide $3.00; Additional stained slide from same block $2.00, unstained $1.00 Frozen: Unstained section: 3:00/slide, section on same slide $1.00/section Floating frozen section: mounted & coverslip $2.00/slide User cuts their own cut their own sections: $20.00 per hour The service provided thus far has been the investigators personnel use of the facility in the preparation and cutting of their specimens. User hours: 280. The core began full service on February 1, 2007. Electron Microscopy Lab Woo-Kuen Lo, Leader Lawrence Brako, Electron Microscope Technician. Established in 1985, the RCMI EM Core facility provides complete scanning and transmission services, from specimen fixation/dehydration/critical point drying/ embedding/polymerization/ block trimming/sectioning/slide-making/formvar-coating, to examination and documentation of prepared samples utilizing JEOL 820 scanning and JEOL 1200EX transmission electron microscopes. Service Charges $150.00 per sample preparation, $2.50 per TEM negative, $5.00 per SEM polaroid. This includes: a) Complete preparation of samples (i.e. fixation, dehydration, critical point drying, sputter coating, embedding, polymerization, knife-making, trimming, H/E slide-preparation, sectioning, formvar-coated grid preparation and subsequent grid- staining...) b) Examination and documentation of prepared specimens utilizing Transmission and/or Scanning Electron Microscope. c) EM negatives scanned to CD. The generated revenue covers cost of basic EM materials, chemicals, Polaroid film, EM film negatives and general supplies, diamond knife replacement ($3000.00), equipment repairs and maintenance (eg. critical point dryer overhaul, $1500.00; Ultramicrotome service, $1500.00) and small equipment purchase Year # of EM negatives processed # of Users 2006 420 14 a) The total number of biological samples processed in an average year is between 80-120. b) Average time per biological sample preparation (including Fixation, Dehydration, Critical Point Drying, Embedding, Polymerization, Knife-making, Block Trimming, Sectioning, Formvar-Coat preparations, Gold/Palladium sputtering, Grid-staining) amounts to 45 Hours (though actual turn-around time is approximately 7-10 days). c) Average microscope time per user ranges from 10-40 hours per year, each session lasting about 2 hours which allows more users access to the microscopes in the same day. Imaging Lab Xieubao Yao, leader The Image Analysis Core (IAF) presently consists 3 primary pieces of equipment: a newly acquired Leica TCS SP5 confocal microscope, a Tecan plate reader capable of fluorescent, luminescent and absorbance measurements, and a Zeiss Axioplan upright fluorescent microscope with attached Spot Insight digital camera and software. Also housed in the facility are 2 incubators (1 CO2 , water jacketed), a cell culture hood and a Zeiss Axiovert 25 cell culture microscope to aid in the maintenance of cells while performing experiments in the IAF. Oversight of the facility is provided by Dr. Xuebiao Yao, Director of the IAF and Assoc. Professor in the Dept. of Physiology (5%) and Mr. Andrew Shaw, Lab coordinator (90%). Service Charges: 1. Confocal microscopy: $40 / hr attended; $30 / hr unattended. User must demonstrate proficiency with the equipment before unattended use will be allowed. 2. Tecan Plate Reader: $20 / hr. Average run takes approximately 5 min.; however, time-course experiments will be billed for the entire time that the equipment will be reserved. 3. Zeiss upright fluorescent microscope: $20 / hr.