Flow cytometry is a powerful technology for analyzing and sorting a suspension of microscopic particles at thousands of events per second. The technique utilizes fluorescent probes targeted to cell specific antigens to characterize the physical and/or chemical characteristics of single cells. Fluorescence-activated cell sorting (FACS) coupled with analysis can identify and isolate live cells from a defined phenotype that can be expanded in culture or in animal models and further analyzed.
Flow analyzers are available 24/7 upon completion of the mandatory training, while cell sorters are CIC operated only.
For more information about training, see the Training Information section below.
|Cell sorters||Sorting Rates*||Setup Fee*|
|Thermo Fisher Bigfoot||$90/hr||$50|
|Beckman Coulter MoFlo XDP||$60/hr||$80|
*Minimum charge 30 minutes
|Analyzers||Unassisted rates||Staff-assisted rates|
|Beckman Coulter CyAn ADP||$40/hr||$60/hr|
|BD LSRFortessa X-20||$55/hr||$75/hr|
Authorized users will have access to the instruments 24/7 following completion of the training and approval from a CIC staff member. Instruments can be reserved on the online calendar using the credentials provided by a CIC staff member.
The Cytek Aurora spectral analyzer is equipped with four lasers (405, 488, 561, and 640 nm excitation wavelengths) and three scatter channels. The highly sensitive violet side scatter detector allows for the analysis of particles approaching the 100 nm size. The detectors in this instrument capture a fluorescence signature along a continuous spectrum and generate a spectral signature for up to 48 fluorescent channels. The new intuitive SpectroFlo software offers streamline assay setup, live data unmixing, auto-fluorescence extraction, and FCS 3.1 file export. The configuration of this instrument is similar to the Thermo Fisher Bigfoot cell sorter, minus the ultraviolet laser, making the transition between running samples on the Aurora to sorting on the Bigfoot relatively easy.
Thermo Fisher Bigfoot
The Thermo Fisher Bigfoot full-spectrum sorter is equipped with five lasers (355, 405, 488, 561, and 640 nm excitation wavelengths) and a 405 nm side scatter detection for increased resolution of small particles. This spectral sorter measures the entire emission spectra and allows the capture of up to 48 fluorescence channels. The high-speed jet-in-air system is well suited for sorting fragile cells and can achieve sort rates up to 70,000 events per second. Up to six individual samples can be loaded at one time, and this is accompanied by built-in sample agitation and temperature control. Standard six-way sorting can be carried out into a variety of output holders ranging from 1.5 to 50 mL tubes, and multi-well plates up to 1536 wells. The configuration of this sorter is similar to the Cytek Aurora, with the addition of an ultraviolet laser, facilitating the transition from analysis with the Aurora to sorting with the Bigfoot. The Sasquatch software offers the ability to analyze/sort samples using spectral mode or compensation mode.
BD LSRFortessa X-20
The BD LSRFortessa X-20 is configured with four lasers (488 nm, 640 nm, 405 nm, and 552 nm) capable of detecting up to 15 colors and 17 parameters. The dual forward scatter detection system includes a highly sensitive forward scatter PMT, allowing for the recognition of particles as small as 200 nm. The instrument is equipped with the FACSDiva 8.0 Software.
Beckman Coulter MoFlo XDP
The MoFlo XDP is a high-speed jet-in-air cell sorter with the ability to sort up to 4 populations simultaneously. The system is equipped with two lasers (488 nm and 635 nm) and can detect up to seven colors. This instrument can achieve sort rates up to 70,000 events per second with high viability, and is well suited for purifying fragile cell types as well as rare populations present at <1% of the starting population. Cells with defined characteristics can be deposited directly into 96 well plates with precision, facilitating downstream applications requiring one or more cells per well. The configuration of this instrument is similar to that of the CyAn ADP, making it possible to transition easily from analysis on the CyAn ADP to sorting on the MoFlo XDP.
Beckman Coulter CyAn ADP
The CyAn ADP cytometer is an easy to use instrument configured with two lasers (488 nm and 635 nm) and the ability to detect up to seven colors. Patented technology enables high acquisition rates of up to 70,000 events per second, ideal for analyzing rare event populations. Compensation modes include both manual and automatic options. The configuration of this instrument is similar to that of the MoFlo XDP, making it possible to transition easily from analysis on the CyAn ADP to sorting on the MoFlo XDP. The instrument is equipped with Summit software, and the data can be analyzed using a free desktop version of this software or analyzed using other third party software programs.
The CIC staff will provide training to investigators wanting to learn more about flow cytometry and how to operate the flow analyzers. Training is mandatory even if the investigator has previous experience in flow cytometry or using other flow analyzers. The mandatory training sessions are offered by appointment at a rate of $40.00 per training session in addition to the fee for using the instrument during the training. Training is typically completed in two hands-on sessions of 60-90 minutes each. The first training covers a theoretical lesson of flow cytometry and the policies of the CIC facility. The second part involves actual time using the instrument and a guided run through practice experiment. Investigators must provide samples appropriate for the analyzer for the second training. Following the instrument training, a second session with a CIC operator may be needed until the user can be approved to use the instrument independently. Once approved, a username and password will be provided for instrument access and the online reservation calendar.
Training Guide (Please print, review and bring to your first training session.)
- Flow cytometer reservations
- Troubleshooting on the FACScan
- FACScan CIC customer guide
- FACSCalibur CIC customer guide
- BD flow PowerPoint
- Molecular probes spectra viewer
- Fluorochrome absorbption and emission spectra
- CyAn fluorochrome guide
- CyAn CIC customer guide
- BD Fluorochrome reference & specification chart
- BD's spectrum viewer
Flow cytometry protocols
- 7-AAD viability staining
- CBA setup
- CBA users guide
- Cell cycle with PI & BrdU
- Cell cycle with PI
- Cell cycle basics
- Immune function analysis techniques
- Immunofluorescent staining of leukocytes
- Tracking cell proliferation with CFSE
- Cell cycle analysis by flow