The Albert Einstein College of Medicine (AECOM) Flow Cytometry Core Facility is a Cancer Center-subsidized shared resource. Its function is to provide high quality, cost effective state-of-the-art flow cytometry and multiparameter cell sorting instrumentation and associated expertise and services to all investigators at the college. The facility permits maximum and efficient utilization of the equipment and serves to enhance the quality and scope of scientific research performed at Albert Einstein.
The high cost of the instrumentation in the core and the need for the specialization of skilled personnel mandated the availability of a shared resource for flow cytometry at the medical school and a flow cytometry core facility was incepted in 1976 when a Becton Dickinson FACS II was purchased using a National Science Foundation equipment grant. It is interesting to note that the first commercially available cell sorter, which was a Becton Dickinson FACS, was introduced only 2 years earlier.
The Flow Cytometry Core Facility is equipped with 6 flow cytometers that are categorized into 2 types:
Bench top analyzer flow cytometers that are operated by the investigators themselves and High-speed cell sorter flow cytometers that are operated by the core facility's personnel as a service to investigators only.
Core Facility Bench Top Analyzer Flow Cytometers:
There are 5 bench top analyzer flow cytometers in the core that are manufactured by Becton Dickinson Immunocytometry Systems, San Jose, California, and consist of 1 upgraded, 4-laser FACScalibur, 1 upgraded dual-laser FACScan, a dual-laser FACSCantoll, a four-laser LSRII and a five-laser LSRII. Know that FACS™ is an acronym for Fluorescence-Activated Cell Sorting and is Becton Dickinson's registered trademark.
Core Facility High-Speed Cell Sorter Flow Cytometers:
There are 3 high-speed cell flow cytometers in the core facility that consist of 3 different models by 2 manufacturers. The facility is equipped with a DakoCytomation MoFlo and a Dako (formerly DakoCytomation) MoFlo XDP. Dako is based in Fort Collins, Colorado. The 3rd sorter in the facility is a Becton Dickinson FACSAria.
The primary function of the high-speed cell sorter flow cytometers are for physically sorting cells or particles of interest and for analysis requirements that cannot be met on the bench top analysis flow cytometers in the facility. For example, the sorter flow cytometers are the only flow cytometers in the facility that are equipped with cyan, green, yellow, orange or far red laser lines, consequently, investigators requiring access to these excitation lines are granted access to the sorter flow cytometers regardless if their goal is to physically sort cells (particles) of interest, otherwise, as stated previously, the primary function of the sorter flow cytometers is for physically sorting cells or particles of interest.
Unlike the bench top analyzer flow cytometers, which are operated by the investigators themselves, sorter operation is provided as a service. This is because the instruments are much more complex than the bench top analyzers and consequently require much more experience and training to achieve proper function. Also, because of the open design of the MoFlos, which permits great flexibility in experimental design, their components are more easily damaged and they are expensive to replace.
Because the sorters use a stream-in-air sorting method, they aerosolize the samples. Consequently, the sorters cannot be used for sorting hazardous samples. Non-hazardous living cells can be sorted and recovered in sterile form for subsequent culture or for in vitro functional studies. Accommodations can be made for AECOM investigators requiring sorting of hazardous specimens off site - please inquire in advance.
Miltenyi Biotec SuperMACS manual magnetic antibody bead separator, for large-scale (bulk) cell sorting, available to trained users with online booking.
The SuperMACS allows you to manually separate more than 1011 total cells in one step via a single-use disposable column. Those wishing to achieve the highest purity or are separating rare cells should consider using two columns successively per separation. The SuperMACS is designed for gentle and ultra high speed positive selection as well as depletion. The SuperMACS operates inside a laminar flow hood to insure a sterile environment.
The SuperMACS is compatible with almost any direct and indirect MACS® reagents for the isolation of virtually any cell type. Cells that can be easily and efficiently isolated by using the SuperMACS include human, rat, and mouse cells or cells from any other species, for example CD34+ progenitor cells, CD3+, CD4+, CD8+, CD14+, CD15+, CD56+ cells from human whole blood, cytokine-secreting cells, dendritic cells and disseminated tumor cells, among others. You only have to magnetically label the cells and run the cells over the column.
The SuperMACS is completely compatible with flow cytometry. Fluorescent and magnetic labeling of cells can be performed simultaneously. After SuperMACS sorting, cells are immediately ready for flow cytometric analysis. MACS MicroBeads, for example anti-FITC or anti-PE MicroBeads, are added to FITC or PE stained cells. Then the cells are sorted on the SuperMACS and the positive and negative fractions are collected 12 x 75 millimeter polypropelyne test tubes for convenient flow cytometer processing.
Becton Dickinson FACSCalibur: DxP10 FACSCalibur upgrade for our FACSCalibur make the cytometer capable of 10 channels of fluorescent detection excited by four lasers (488/561/637/407). Electronic signal processing is also upgraded from the original analog paths to full digital processing. Acquisition control has the added benefit of the long trusted analysis software, FlowJo, in a "Collectors" edition suite
The core is equipped with 1 upgraded FACScan flow cytometer that is equipped with a low-power (15mW) argon laser that emits blue light at 488 nanometers as well as a low-power (30mW) helium-neon diode laser that emits red light at 633 nanometers. The core's FACScan is upgraded from the factory's original configuration that has only 1 laser and 3 fluorescent detectors. It has two detectors for laser-light scatter and 4, not 3, detectors for fluorescence in the green, orange and red/dark red regions of the color spectrum. The detector array permits the use of a great number of fluorophores, allowing for up to 7 parameters for every particle interrogated (time is also a parameter). The optical filters on the FACScan are not interchangeable (each fluorescence detector "sees" on a single fluorescence color) so the choices of fluorophores are similarly constrained, however, a tremendous number of fluorophores can be detected with it's configuration.
Both FACScans are equipped with pulse processing for cell doublet discrimination and have an optional 20-liter cubitainer system that has 5 times the buffer and waste capacity of the factory system.
Becton Dickinson FACS CantoII: The BD FACSCanto II features many innovations for both clinical and research labs, including a true fixed-alignment flow cell to minimize startup time and improve reproducibility. With High Throughput Sampler (HTS) option available for this flow cytometer. The HTS provides fully automated and rapid sample acquisition from either a 96- or 384-well microtiter plate.
AECOM Flow Cytometry Core has not received any reviews.
AECOM Flow Cytometry Core has not received any endorsements.