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Flow Cytometry

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Equipment

Coulter Cytomics FC500 flow cytometers: These are user-friendly analytical instruments with one air-cooled argon laser and one air-cooled helium-neon laser that permit simultaneous measurement of five different fluorescent probes. These instruments are used for routine multiparameter analysis of DNA content, cell cycle, immunofluorescence, apoptosis, green fluorescence protein expression, oxidative metabolism and phagocytosis. Two Cytomics FC500 instruments are currently located on the Busch campus, and staffed by Theresa Choi and Joan Dubois. A third Cytomics FC500 is located at CINJ and is staffed by Art Roberts. Schedule the Flow Cytometer at CINJ

Beckman-Coulter MoFlo XDP Cell Sorter: The MoFlo allows rapid separation of large numbers of specific cell populations with high purity, recovery and yield.  It an extremely stable platform allowing for analysis at rates up to 100,000 events/second and sort speeds of up to 70,000 events/second.  In addition, the instrument is capable of simultaneous 4 parameter sorting.  The MoFlo is equipped with 3 solid-state air-cooled lasers allowing for excitation wavelengths of 355, 488 and 633 nm and 9 PMTs making it possible to simultaneously analyze 12 different cell properties. The instrument also has a biosafety containment cabinet and an automatic shut-down mode which will permit sorting of transfected human cells.  In addition, the MoFlo is equipped with the Cyclone® sorting option for single cell cloning or for sort matrix verification.   

Leica TCS SP5 Spectral Confocal Microscope: This is a laser based confocal system that permits measurements of fluorescently labeled tissue sections, individual cells, or particles adhered to matrices over a wide selection of visible wavelengths. The instrument is equipped with 5 air cooled lasers allowing for excitation wavelengths of 405, 458, 476, 488, 496, 514, 561, 594 and 633 nm. Analysis of immunofluorescence as well as functional and biochemical kinetic studies can be performed on viable cell samples. Intracellular distribution of fluorescence can also be quantified. The instrument has extremely high sensitivity and can differentiate between closely emitting chromophores. In addition, the Leica TCS SP5 features digital resolution of 64 megapixels X12 bits per channel with 4 detector channels and can measure real time response of time-critical sequences due to its fast scan speed. The Leica TCS SP5 has a full complement of objectives for collecting high-resolution 3-D images. Laser scanning confocal microscopy will offer CINJ members improved resolution and dramatically improved rejection of out-of-focus light in comparison to conventional epifluorescence microscopy. Optical sections of 1 to 5 mm can be collected at depths up to 150 mm into a specimen. The image is acquired by illuminating each pixel in series by raster-scanning an exciting laser beam over the specimen; a confocal aperture is used to collect the light emitted from only the excited pixel. The Leica confocal microscope is currently located on the Busch campus, and staffed by Dr. Carol Gardner.

Xenogen IVIS® imaging system: This instrument provides investigators with the capability to perform real-time in vivo imaging of small animals. The ultra-sensitive CCD cameras, hardware and software allow detection of emitted photons and the creation of images and data files. The technology makes possible the tracking of biological activity in real-time, at the molecular level. This real-time in vivo imaging utilizes luciferase to visualize markers of interest. By imaging the whole animal at many individual time points, researchers can better understand disease pathology, pharmacokinetics, response to therapy, and other contextual or spatial aspects of the processes taking place in the living animal. The Xenogen IVIS is currently located on the Busch campus, and staffed by Dr. Vasanthi Sunil.