Flow Cytometry Core Service Request
The BD FACS Calibur cytometer has two lasers, 488 and 633, 2 scatter detectors, and 4 PMTs capable of 4 color analysis with the two excitation wavelengths mentioned. 3 color manual compensation is available. The Calibur uses BD Cell Quest Pro acquisition and analysis software. For additional information on the use of the Calibur go to the Calibur User Guidelines for step by step instructions or Calibur Quick Notes for a condensed operational guide.
The flow core has three BD LSR II cytometers available for your use. Starting with the Dodecapus and progressing to either the Behemoth or the Octopus each of these cytometers offers increased capacity. Additionally, since all three cytometers utilize DIVA acquisition software there is a level of redundancy across all three machines assuring that regardless of schedule conflicts or machine downtime one should have access to one machine or another at any time. All LSR’s have automatic compensation capability. For additional information on the use of the LSR’s go to the Filter Configurations and/or LSR User Guidelines for in depth, step by step instructions. You can also access the LSR Quick Notes for a condensed version on how to operate the LSR II.
FACSVantage II DiVa
Fluorescence Activated Cell Sorting (FACS) is available to our customers and performed on the Becton Dickinson FACSVantage cell sorter. Having 3 lasers (488, 633 and UV), 2 scatter detectors and 8 fluorescent detectors, the Vantage is capable of a wide variety of cell sorting. The Vantage can be configured for 2-way or 4-way sorting as well as macro sorting, high speed sorting, and single cell sorting into 96 well plates, which is perfect for cloning or single cell analysis. In addition the Vantage is equipped with a sample cooling system should sort and collection temperature be an issue for your samples.
A flow cytometer sorter makes it possible to isolate highly purified populations of particles having distinct characteristics. These particles are not limited to cells, but may be chromosomes, microorganisms, microbeads, etc. Any parameter that is measurable on a flow cytometer can be used as a sorting criteria. DiVa software includes width and height measurements for all PMTs, so doublet discrimination and DNA analysis are available. DiVa software performs compensation on all channels, including cross beam compensation.
The Vantage works by rapidly vibrating a crystal connected to a flow cell, causing a stream of cells to break into uniform droplets. As each droplet separates from the stream it can be independently charged and subsequently deflected past a charged plate into a receptacle. The instrument can sort 4 independent populations at a time with a fifth stream containing unwanted cells. The Vantage utilizes BD DiVa acquisition and analysis software. Data output is through the USB drive.
For additional information on preparing cells for sorting in order to optimize your results or to view the filter configuration on the Vantage go to the Sort Preparation Page or the Filter Configurations PDF.