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
Vantoo DiVa (BD Digital Vantage): Vantoo is a 7 color 2 laser sorter upgraded with digital electronics. This sorter is capable of high sorting speeds, simultaneous sorting of 4 distinct populations at once, and real time full matrix compensation. Its current configuration is approved for sorting BSL-2 samples.
Model: BD Digital Vantage
Room: CCSR 3232
Sort: yes Clone: yes
cells/sec (max): 10,000 (35 psi), 4,000 (15 psi)
BSL-2: yes
Colors: 7
Excitations (nm): 488, 594
Aria
Aria: Aria is a 3 laser 11 color fixed alignment sorter. It's basic capabilities are similar to Vantoo, with a slightly different laser configuration and an easier-to-use software interface. This instrument is approved for sorting BSL-2 samples. Model: BD Aria  II
Room: B016C
Sort: yes Clone: yes Index Sort: yes
cells/sec (max): 25,000
BSL-2: yes
Colors: 11
Excitations (nm): 488, 635, 405
Falstaff
Falstaff: Occupying a larger footprint than Aria, Falstaff is a BD Special Order sorter, with 5 lasers and a 17 color detector configutation. The additional laser lines allow use of fluorochromes and fluorescent dyes that cannot be detected on 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 Clone: yes Index Sort: yes
cells/sec (max): 25,000
BSL-2: yes
Colors: 17
Excitations (nm): 488, 532,635, 405, 355.
Aida Aida: "Aida" is a 4 laser Aria II sorter similar to "Falstaff". It does not have a UV laser, and has a 561 laser instead of a 532 laser. It can detect up to 13 colors at once and is approved for BSL-2 sorting. It was purchased by the Department of Medicine; some access and biological restrictions apply. Model: BD Aria  II
Room: B016B
Sort: yes Clone: yes Index Sort: yes
cells/sec (max):25,000
BSL-2: yes
Colors: 13
Excitations (nm):488, 561,635, 405.
VerdiVerdi is a 3 laser Aria II sorter. It does not have a violet laser, and can only detect up to 7 colors at once. Verdi is approved for BSL-2 sorting.  Model:  BD Aria II
Room: B016C
Sort: Yes Clone: Yes Index Sort: Yes
cells/sec (max): 25,000
BSL-2: yes
Colors: 7
Excitations (nm): 488, 561, 640.
PICIPICI is a 5 laser Aria Fusion sorter that was bought with Parker Institute for Cancer Immunotherapy funds. It can detect up to 17 colors at once, and is approved for BSL-2 sorting.  Model: BD Aria Fusion  II
Room: B019
Sort: yes Clone: yes Index Sort: yes
cells/sec (max): 25,000
BSL-2: yes
Colors: 17
Excitations (nm): 488, 561, 640, 405, 355.
Carmen (BD InFlux): Carmen is a 7 laser 22 color sorter. It can sort up to 4 populations at once, into vessels ranging from 1.5 mL to 50 mL tubes, to 6-well to 384 well plates, to microscope slides, to unique customized configurations. The multiple lasers allows the measurement of any fluorochrome or fluorescent protein that is commonly available.  Carmen offers full matrix compensation, and is approved for sorting BSL-2 samples.  Model: BD Influx Special Order
Room: B015A
Sort: yes  Clone: yes Index sort: yes
cells/sec (max): 25,000
BSL-2: Yes
Colors: 22 available; 16 max at one time
Excitations (nm): 375, 405, 457, 488, 532, 561, 640
\ Satchmo (BD FACSJazz) is a simple two-laser (488 nm and 640 nm) sorter. It is easy to operate, and does fast tray sorting, as well as index sorting. It uses the same plyproplyne sample tubes as Carmen. Model: BD FACSJazz Sorter
Room: B016B
Sort: yes  Clone: yes Index sort: yes
cells/sec (max): 15,000
BSL-2: Yes
Colors: 6 available; 4 excited by 488nm, 2 excited by 640 nm
Mikado (Sony SH800s) is the most automated sorter that the Facility has. It is simple to use, and has tray and idex sorting. It is the best sorter for 384 trays, It has 3 lasers (405 nm, 488nm, 561nm) but is a little limited in fluorochrome choices as these lasers are co-linear, Model: Sony SH800s sorter
Room: B016B
Sort: yes  Clone: yes Index sort: yes
cells/sec (max): 5,000
BSL-2: Yes
Colors: 6 available; choose from 405 nm, 488nm,or 640 nm excitation.
LSR II
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
BSL-2: yes
Colors: 14
Excitations (nm): 488, 532, 635, 405
LSR II.UV: A near relative of the twins above. This analyzer is distinguished by having a UV laser and a heating system for doing Ca+ flux experiments. It also has 4 visible lasers and is  equipped with a 96 well plate reader in addition to a standard 5 ml sample tube station. It can detect up to 14 colors and eperiments which are designed for the other two LSRs should, in general, work on this instrument as well. Violet laser colleciton is limited to 2 PMTs with the addition of the UV laser. This instrument is located in BSL-2 lab space and is approved for analyzing BSL-2 samples.
Model: BD LSR II
Room: B016C
Sort: no
cells/sec (max):25,000
BSL-2: yes
Colors: 14
Excitations (nm):355, 488, 561, 635, 405
Scanford: The FACScan Analyzer was one of the first successful bench top flow cytometers. They just keep going and going. This one was purchased in 1990 and has been customized by Stanford with a pressure gauge and adjustable flow settings, and by Cytek Biosciences with 4 lasers, associated optics, and digital electronics to measure 12 colors at once. This instrument is located in BSL-2 lab space and is approved for analyzing BSL-2 samples. Model: Custom Stanford and Cytek upgraded FACScan
Room: B016C
Sort: no
cells/sec (max):3,000
BSL-2: yes
Colors: 12
Excitations (nm): 488, 405, 561, 640.
Cyrano: The CyTOF2 Mass Cytometer is an analysis instrument capable of measuring dozens of experimental parameters at the single cell level, using metal-ion conjugates and mass measurements to differentiate functional and phenotypic markers.  Such an approach precludes the use of compensation and expands the experimental panel size potential, each factors limiting the expansion of work with traditional flow cytometry.  Sample preparation and processing is quite similar to that used in fluorescence-based cytometry with some quality control considerations unique to atomic mass spectroscopy.  Please contact staff for further details. Model: DVS/Fluidigm CyTOF2
Room: B015A
Cells/sec (max): 500
Parameters: ~100

 

 

      Instrument Specifics

 Facility Name
 Model Room  Sort  Clone  Plate Reader Chiller/Heater maximum cells/second minutes to sort 106 cells BSL-2  Colors  Lasers Excitations

Price/hr no Operator

Aria BD Aria II B016C Yes Yes No chiller 25,000 4 Yes 11 488nm, 635nm,
405nm
$70
Falstaff BD Aria II B019 Yes Yes No both 25,000 4 Yes 17 488nm, 532nm, 635nm, 405nm, 355nm $70
Aida BD Aria II B019 Yes Yes No both 25,000 4 Yes 13 488nm, 561nm, 635nm, 405nm $70
Verdi BD Aria II B016C Yes Yes No chiller 25,000 3 Yes 7

488nm,

561nm,

640 nm

$70
Vantoo BD Digital Vantage CCSR 3232 Yes Yes No chiller 10,000 (35 psi)
4,000 (15 psi)
17
40
Yes

7

488nm, 594nm
$55
Carmen BD Influx Special Order B015A Yes Yes No both  25,000  4 Yes 22 355nm 405nm, 457nm, 488nm, 532nm, 561nm, 640nm $75
Satchmo BD FACSJazz B016B Yes Yes No Chiller 10,000 20 Yes 6 488 nm, 640 nm $50
Mikado Sony SH800s B016B Yes Yes No No 7500 35 Yes 6 405nm,488nm,561 nm $60
LSR II BD LSR II B016A No No Yes none 25,000
N/A Yes 18 488nm, 532nm, 635nm, 405nm $55
LSR II.2 BD LSR II

B016A

No No Yes none 25,000
N/A Yes 18 488nm, 532nm, 635nm, 405nm $55
LSR II.UV BD LSR II B016D No No Yes none 25,000 N/A Yes 14 488nm, 561nm, 635nm, 405nm, 355nm $55
Scanford Custom FACScan B016D No No No none 3,000
 N/A 
Yes 12 488nm, 405 nm, 561nm, 640nm $40
Cyrano DVS CyTOF II B015A No No Yes none 500 N/A Yes ~100 N/A $130

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

The sort 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 is cumbersome 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: 02/10/14