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. DESCRIPTION Cytometry and Cell Sorting Lab - Udai P Singh, Leader Advanced cell sorting and flow cytometry analysis have broad multidisciplinary uses to conduct experiments in many 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 has facilitated access and ensured effective use of the Core's capabilities. The flow cytometry has provided advanced and reliable technical support to MSM researchers as well as others institution using this technology. This technology has also enhanced the research and training environment for undergraduate, graduate and medical students engaged in research at MSM. 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. Electron Microscopy Lab - Woo-Kuen Lo, Leader 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. Imaging Lab - Xieubao Yao, Leader The Image Analysis Core Facility(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: Cytometry and Cell Sorting Lab 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 Service Charges: Cytometry and Cell Sorting Lab 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 Service Charges: Cytometry and Cell Sorting Lab 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 Molecular Histology Lab 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 $5.00; Cut, Stain, Coverslip -1st slide $4.00; 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 Electron Microscopy Lab $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 gridstaining...) b) Examination and documentation of prepared specimens utilizing Transmission and/or Scanning Electron Microscope. c) EM negatives developed and scanned onto CD for investigator. 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 Imaging Lab Confocal microscopy: $40 / hr attended; $30 / hr unattended. User must demonstrate proficiency with the equipment before unattended use will be allowed. Tecan Plate Reader: $20 / hr. Average run takes approximately 5 min.; however, timecourse experiments will be billed for the entire time that the equipment will be reserved. Zeiss upright fluorescent microscope: $20 / hr