Available Instruments & Capabilities

Training is available for Facility users to learn to self-operate analyzers and sorters. Operator support is available for those that would rather have staff assistance in running/sorting samples. For the LSR II analyzers a 96 well plate reader is available.

Available Instruments
	Vantoo DiVa (BD Digital Vantage): Vantoo is a 12 color 3 laser sorter with digital electronics. It is capable of high sorting speeds, can sort 4 populations at once, and can do real time full matrix compensation.  Cloning is supported on Vantoo. It supports UV (355 nm) excitation. This instrument is approved for sorting BSL-2 samples.	Model: BD Digital Vantage Room: B015A Sort: yes cells/sec (max): 10,000 (35 psi)                              4,000 (15 psi) Minutes/106 cells: 17 (35 psi)                                40 (15 psi) BSL-2: yes Colors: 12 Excitations (nm): 488, 355,                      570-595 tunable
	Aria: The Aria is a 3 laser 11 color fixed alignment sorter. It's basic capabilities are similar to Vantoo, but it is easier to learn to use. This instrument is approved for sorting BSL-2 samples.	Model: BD Aria  II Room: B016C Sort: yes cells/sec (max): 25,000 Minutes/106 cells: 4 BSL-2: yes Colors: 11 Excitations (nm): 488, 635, 405
	Falstaff: Falstaff is visually higher and wider than the Aria to accomodate its 5 lasers.  Its detector configutation allows for 17 colors.The addistional laser lines allow use of fluorochromes and fluorescent dyes that cannot be detected on the Aria, including UV excited fluochromes and green laser excited fluorescent proteins. This instrument is approved for sorting BSL-2 samples.	Model: BD Aria  II Room: B016B Sort: yes cells/sec (max): 25,000 Minutes/106 cells: 4 BSL-2: yes Colors: 17 Excitations (nm): 488, 532,635, 405, 355.
	Flipper: Flipper is our custom sorter (Flasher II in semi-retirement). It is a  2 laser 8 color instrument constructed from various parts of different commercial instruments. It has a customized Facstar laser bench, extensively modified optics, and DiVa electronics.  Cloning is not supported on this instrument. The CCSR site is our first experiment at managing remote sites. Access to it outside of the Baxter labs is limited to operator supported use.	Model: Custom Hybrid Room: CCSR 3232 Sort: yes cells/sec (max): 4,000 Minutes/106 cells: 40 BSL-2: no Colors: 8 Excitations (nm): 488,                       570-595 tunable
	Carmen (BD InFlux): Carmen is a 7 laser 22 color sorter. It can sort up to 4 populations at once, into tubes of sizes from Eppies to 50ml, as well as into plates of different sizes. The multiple lasers allows the measurement of any flurochrome or fluorescent protein that is commonly available.  Carment offers full matrix compensation, and is approved for sorting BSL-2 samples. The Facility is collaborating with BD Biosciences on evaluating novel flow applications on this instrument, so we are actively soliciting users who have cytometry applications they have not been able to perform on other existing instruments	Model: BD Influx Special Order Room: B016D Sort: Yes cells/sec (max): 25,000 Minutes/106 cells: 4 BSL-2: Yes Colors: 22 available; 16 max at one time Excitations (nm): 375, 405, 457, 488, 532, 561, 640
	LSR II and LSR II.2: These analyzers are nearly identical twins. They are 4 laser 14 color instruments, each equipped with a 96 well plate reader in addition to a standard 5 ml sample tube station. These instruments are located in BSL-2 lab space and are approved for analyzing BSL-2 samples.	Model: BD LSR II Room: B016A Sort: no cells/sec (max): 25,000 Minutes/106 cells: 4 BSL-2: yes Colors: 14 Excitations (nm): 488, 532, 635, 405
	LSR I: An early version of the LSR analyzer. This has 2 lasers and supports 6 color measurements. This instrument is located in BSL-2 lab space and is approved for analyzing BSL-2 samples.	Model: Custom LSR I Room: B016B Sort: no cells/sec (max): 4,000 Minutes/106 cells: 40 BSL-2: yes Colors: 6 Excitations (nm): 488, 635
	Scanford: FACScan Analyzer. One of the first successful bench top flow cytometers. These just keep going and going. One laser and 3 colors. This instrument is located in BSL-2 lab space and is approved for analyzing BSL-2 samples.	Model: Custom FACScan Room: B016C Sort: no cells/sec (max): 2,000 Minutes/106 cells: 80 BSL-2: yes Colors: 3 Excitations (nm): 488

The above rates  for sorting are for the smallest nozzles the SSFF supports; 70 micron on the Aria, 80 micron on Vantoo and Flipper, and 75 micron on Carmen. Maximum sort rates will be slower for larger sized nozzles on these instruments.

The rates above do not distinguish between cells and debris. If you are sorting for culture or injection, then debris in sorted sample probably would not affect your results. Thus a high threshold could be set and most of the debris would be ignored. If you are sorting to do a protein or nucleic acid assay (e.g. PCR), then debris could adversely affect your results. Thus a low threshold would be advantageous, and the cell rate would be reduced by the amount of debris measured.

For Rare Event Sorting recovery (yield) and throughput will be enhanced by doing a two-pass sort. This can be useful when the desired population frequency is as high as 5%. For more information or to discuss further please contact Facility Staff.

A third consideration is the frequency of the desired population. The time estimates do not account for the frequency of aborts due to coincident events. Sort aborts are inversely proportional to the frequency of the desired sorted population; lower frequency results in a higher abort rate. For very infrequent populations, the event rate must be slowed down considerably to maintain an acceptable yield. If an infrequent population, on the order of 1%, is desired from a large number of cells, 10 million or so, then a two pass sort will be more efficient. The first pass will enrich the population to about 30%. When spun down and resuspended in 1/2 ml, a purity sort at the event rates above will be possible, giving high yield and purity in an acceptable amount of time. Please let Facility staff know if this situation may apply to you.

Ask Us!

It would be difficult to list all the possible fluorochrome/fluorescent protein combinations for each instrument. If you have not used the Facility before, or are planning a new experiment, please check with our staff to select the optimal instrument/fluorochrome combination.

last update: 08/31/09