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Kathy Q. Cai
Last Updated: More than One Year ago
The UNMC Cell Analysis Facility provides flow cytometryservices to investigators on and off the UNMC campus on a fee for service basis.
Services include:
•Immunophenotyping
•cell cycle and DNA content analysis
•protein expression and bead based assays for signal transductionmolecules
•soluble proteins cytokines and growth factors, as well as DNA fragments
Instrumentation includes: 1) three B-D FACSCaliburdual-laser cytometerscapable of four color, 8 parameter analysis, 2) a B-D LSR II four-laser cytometercapable of 13 color,
15 parameter analysis, 3) a B-D FACSArraydual-laser bioanalyzercapable of four color, plate-based multiplex assays, 4) A B-D FACSAriafour-laser, high speed digital cell sorter capable of 15 color, 17 parameter analysis, 5) a B-D FACSVantageDiVathree-laser, high speed cell sorter capable of 6 color, 10 parameter analysis, 6) a Becton Dickinson FACStarPlusthree-laser, low pressure cell sorter capable of 4 color, 6 parameter analysis, and 7) a MiltenyiAutoMACsautomated magnetic cell separation system.
LCM is equipped with two LCM systems (PixCell II® and Veritas™, Molecular Devices, Sunnywale, CA), which are available for a wide range of applications, including the LCM of specific immunostained or fluorescent cells. Viability of the IR laser-enabled LCM technique has been shown for a variety of different cell types. The LCM procedure is remarkably simple and robust. The method is extraordinarily gentle and ideal for microdissection of single cells or small numbers of cells. LCM system utilizes a laser microbeam that melts a thermoplastic membrane that sticks to the selected cells, which can then be lifted and secured in a microfuge tube containing the appropriate extraction solutions. The transferred tissue on the film retains its original morphology, thereby allowing microscopic verification of the specificity of the captured material. Using this strategy, a single small cluster up to thousands of cells can be procured safely and without contamination. The Agilent 2100 Bioanalyzer and Nanodrop ND1000 monitor the quality and quantity of extracted RNA, DNA, as well as proteins from the LCM sample.