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Provides technical expertise and access to equipment for CHLA investigators who use analytical and sorting flow cytometry. Core scientists are available to perform sorts, and to train and oversee scientists with analysis. A wide variety of flow cytometric methods are used. New methods can be developed and established with investigators.
For data acquired with 2nd floor instrumentation, analysis stations are available for your use in SRT203, SRT205 and SRT206 for free-of-charge data analysis. Software packages currently available include: CellQuest (5.x), FlowJo-Mac (v8.x), Flow Jo-PC (v7.x), VenturiOne (v3), and Modfit (v3)
The DiVa FACS Vantage SE (BD) has three lasers, a 488 nm Laser, a 635 nm, and the Innova I-302 tunable krypton laser with UV, Violet, Green*, Yellow or Red line capability. The Vantage collects up to 8 colors: 4 parameters (laser 1), two parameters (laser 2) and two parameters (laser 3). This is in addition to two scatter parameters as well as derived parameters. The optical system is operator-aligned and optimized daily for specific experimental needs. Designed for flexibility, many specialized assays including Calcium Flux and side population, are performed on the DiVa FACSVantage SE. Recently upgraded with a stand-alone air compressor, the DiVa FACSVantage is capable of ultra slow speed to high speed cell sorting. It can deliver four populations simultaneously to a purity frequently in excess of 99%. Diva (v5.x) is the software driving the instrument.
*new optics have been installed for this capability
The FACSAria-I (BD) instrument is equipped with three lasers, capable of detecting up to 13 colors, and is a high-speed cell sorter. The Aria-I is configured with BD Biosciences’ patented octagon/trigon optical bench and new-age flow cell, providing fixed alignment with stream in air sorting. Delivering four populations simultaneously to a purity frequently in excess of 99%, the FACSArias have been surpassing expectations. The Aria-I laser bench holds solid-state lasers, 488nm, 633nm, and 405nm, which are delivered to the flow cell via fiber optic cabling. Diva software (v6.x) is the software driving the instrument.
The four-color FACSCalibur (BD) analyzer is a two-laser instrument, capable of collecting a maximum of eight parameters: two scatter, four colors (green, yellow-orange, red and a second red), plus two derived parameters. CellQuest Pro (v5.x) is the software driving this instrument.
The three-color FACScan (BD) analyzer is a one-laser instrument, capable of collecting a maximum of seven parameters: two scatter, three colors (green, yellow-orange and red), plus two derived parameters. CellQuest Pro (v5.x) is the software driving this instrument.
The Imagestream 100 (Amnis) is the newest addition to our cytometer line. This novel instrument is the first imaging flow cytometer, encouraging you to “Think outside the dot”. Loaded with three lasers (488nm, 633nm, 405nm) it will capture six (6) digital images (spatial resolution: 0.75NA, 0.5μm pixel size) of each cell, measuring morphology, brightfield and up to 4 parameters of fluorescence. The precisely controlled fluidics 19 positions the cells in the plane of focus as they flow through the system smoothly and without tumbling. The spectral decomposition element splits the cell imagery into six spectral bands, one for each CCD camera. The unique time-delay integration tracks the motion of the cells and increases the signal 1,000 fold before image is recorded. Inspire (v3.x) software in used for data acquisition and IDEAS (v3.x) software is used for data analysis.
Contact us for further information.
Contact us for further information.
Flow Cytometry is a technology used to take fluorescent measurements of special fluorochromes bound to or made by various cells as they are travel in a single file line while confined by a laminar fluid. Common applications involve immunolabeling of extracellular or intracellular proteins with fluorochrome-conjugated antibodies, reactive dyes, or nucleic acid dyes. Excitation of the fluorochromes is achieved with lasers. Data acquisition is achieved with specialized computer software directly interfaced with the cytometer. Listed alphabetically are applications supported by instrumentation at the Saban Research Institute: Absolute Counts, Apoptosis, Calcium Flux, Co-localization*, Cytometic Bead Assay (CBA) for up to 30 soluble cytokines or proteins per sample, DNA Cell Cycle Analysis, Flow-FISH for telomere length, Immunophenotyping for Surface, Intracellular and Nuclear proteins, Live cell sorting, PhosFlow, Rare Event Detection/Sorting, Reporter Molecules, Side Population, Single-cell Cloning, Single Color Apoptosis*, Sizing, Spot Counting for viral load*, Subcellular compartmentalization*, Synapse activity*,Translocation*, and Viability.
*denotes applications specific to the Imagestream 100.
Childrens Hospital Los Angeles
4650 Sunset Blvd
Los Angeles, CA 90027
Smith Research Tower, 2nd Floor
Hisham Abdel-Azim, MD
Director
Telephone: (323) 361-5664
Fax: (323)906-8193
Email: habdelazim@chla.usc.edu
Lora Barsky
Manager
Telephone: (323) 361-5935
Email: LBarsky@chla.usc.edu
Smith Research Tower, 3rd Floor
Michael Sheard, PhD
Director
Telephone: (323) 361-2297
Email: msheard@chla.usc.edu
Jakie LIn
Sort Operator
Telephone: (323) 361-5455
Email: TSLin@chla.usc.edu
Smith Research Tower, 2nd Floor - Contact Lora Barsky (LBarsky@chla.usc.edu)
Smith Research Tower, 3rd Floor - Contact Jackie Lin (TSLin@chla.usc.edu)