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 Molecular Histology Lab  John Patrickson, Leader This core unit was established in 2007 under the leadership of Dr. John Patrickson. Provides 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 (IAF) presently consists 4 primary pieces of equipment: a newly acquired DeltaVision Core deconvolution microscope, a 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 Professor in the Dept. of Physiology (5%) and Mr. Andrew Shaw, Lab coordinator (90%). Service Charges: 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 grid-staining...) 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, 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 Deconvolution microscopy $40 / hr. 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, time-course experiments will be billed for the entire time that the equipment will be reserved. Zeiss upright fluorescent microscope: $20 / hr.