Flow Cytometry Core Facilities and Equipment

Our Facilities and Equipment

Fluorescent Activated Cell Sorters

Beckman Coulter MoFlo AstriosEQ 4 laser system: A high speed cell sorter (up to 70,000 cells/second) and analyzer (up to 100,000 cells/second), with four fixed lasers (405nm, 488nm, 561nm, and 640nm). The AstriosEQ is capable of acquiring 28 parameters simultaneously. In addition, it contains two forward scatter parameters and three side scatter parameters, allowing for analysis of extracellular vesicles. Two-, Four-, and Six-way sorting is available. Sorting can be done into tubes of various sizes, plates and custom slides. Single cell sorting into plates is also available.

Operating software for the MoFlo AstriosEQ is the Beckman Coulter Summit program, version 6.3. Operation of this instrument requires a dedicated technician to align lasers, maintain the instrument and to assure its reliable and effective utilization.

Beckman Coulter MoFlo AstriosEQ 7 laser system: A high speed cell sorter (up to 70,000 cells/second) and analyzer (up to 100,000 cells/second), with six fixed lasers (405nm, 488nm, 532nm, 561nm, 592nm, and 640nm) and one spatially separated UV (355nm). The AstriosEQ is capable of acquiring 32 parameters simultaneously. In addition, it contains two forward scatter parameters and five side scatter parameters, allowing for analysis of extracellular vesicles. Two-, Four-, and Six-way sorting is available. Sorting can be done into tubes of various sizes, plates and custom slides. Single cell sorting into plates is also available.

Becton Dickinson SORP FACSAria II: A special order five laser cell sorters (up to 22,000 cells/second) and analyzer (up to 70,000 cells/second) equipped with a solid state 50mW 488nm laser, JDS Uniphase 40mW 640nm HeNe laser, Coherent Compass 40mW 561nm laser (allowing for the acquisition, analysis and sorting of a variety of fluorescent proteins), Lightwave Mode Lock 60mW 355nm UV laser, and Coherent VioFlame 50mW 405nm violet laser. Besides the forward and side scatter parameters, the instrument is equipped for detection of four fluorescent parameters from the 488nm laser, three parameters from the 640nm laser, five parameters from the 561nm laser, three parameters from the violet laser and three parameters from the UV laser. Sorting can be done into tubes of various sizes, plates and custom slides. Single cell sorting into plates is also available.

Operating software for the FACSAria II is the Becton Dickinson Diva software, version 6.11, operating on an HP xw4600 Core 2 duo. Operation of this instrument requires a dedicated technician to maintain the instrument and to assure its reliable and effective utilization.

Flow Cytometry Analyzers

Cytek Aurora: A spectral flow cytometer that has the 4 laser system (16V-14B-10YG-8R) which offers up to 40 fluorescent parameters. The Aurora has state-of-the-art optics that allow for excellent sensitivity and resolution without needing filter changes. The flat top beam profile lasers are 405 nm is 100mW (violet detector), 488nm is 50mW (blue detector), 561nm is 50mW (yellow-green detector), and the 640nm is 80mW (red detector). The FSC is based off the 488nm with two SSC, one VSSC off the 405nm and the other BSSC off the 488nm. Sampling rates are low 15uL/min, medium 30 uL/min, high 60uL/min and a plate high throughput at 100 uL/min. Does require a separate end analysis program (FlowJo, FCSexpress, ect.). This machine excels in high complexity panels for rare and dim populations. Please make sure on the weekends that QC has been run before doing your experiment.

Violet detector module	16 channels unevenly spaced bandwidth from 420-829 nm
Blue detector module	14 channels unevenly spaced bandwidth from 498-829 nm
Yellow-Green detector module	10 channels unevenly spaced bandwidth from 567-829 nm
Red detector module	8 channels unevenly spaced bandwidth from 652-829 nm

Two Beckman Coulter Gallios: The Gallios is a three laser benchtop analyzer equipped with a 22mW Argon 488nm laser, 25mW 638 laser, and a 40mW 405nm laser. All are solid state diodes which are spatially separated at 125um. Alignment is software controlled using Cyto System Mgmt software and can be controlled remotely. This allows for detection of five fluorescent parameters from the 488nm laser, three parameters from the 638nm laser, and two parameters from the 405nm laser in a proprietary "Boulevard" configuration for greater fluorescent resolution. With multiple measurements and angles of Forward Scatter and a gel coupled pickup filter lens assembly, the Gallios can resolve 0.4um diameter particles from background and maximize detection up to 40um. This instrument is equipped with a Multi-tube Carousel Loader system to automatically analyze multiple samples. Software for acquisition and analysis is the Beckman Coulter CXP, running on an Intel Pentium Core 2 Duo processor. Individual research personnel, following requisite training from the Flow Cytometry Core staff, may independently use and operate this instrument. It is available to trained users for full use even in off hours, including evenings, nights, and weekends. Access to the instrument during off hours is controlled by a coded electronic card access system, which limits access to approved users only. (Please note that support for this instrument will be ending in December 2020.)

Cyntellect Celigo: The Celigo adherent cell cytometer allows in situ brightfield and fluorescent analysis of adherent cells. Cell number, density, area density, diameter, area, and perimeter can all be measured. Using LEDs, the system can obtain data for three fluorescent channels (ex 377nm/em 447nm, ex 483nm/em 536nm, and ex 531nm/em 629nm) and 1 brightfield channel. 384, 96, 24, 12, 6, and 1536 well plates can be analyzed; as well as, T-25 and T-75 flasks. Software for acquisition and analysis is the Cyntellect Celigo software version 1.3, running on a Dell Precision T3500. Individual research personnel, following requisite training from the Flow Cytometry Core staff, may independently use and operate this instrument. It is available to trained users for full use even in off hours, including evenings, nights, and weekends. Access to the instrument during off hours is controlled by a coded electronic card access system, which limits access to approved users only.

Two Beckman Coulter CytoFLEX LX: The instrument has the capacity for 23 parameters, including 21 for fluorescence detection. The fully activated instrument includes three fluorescent channels from the 355 nm (UV) laser, five from the 405 nm (Violet) laser, three from the 488 nm (Blue) laser, five from the 561 nm (Yellow Green) laser, three from the 638 nm (Red) laser, and two from the 808 nm (Infrared) laser. Option to configure Avalanche Photo Diode detector array to collect side scatter signal from Violet (405 nm) laser. The configured channel (VSSC) can be used to better resolve nanoparticles. 96-well Standard Flat, U and V bottom plates, and 96-deep well plates.

Operating software for the CytoFLEX LX is the Beckman Coulter CytExpert program, version 2.3.1 Operation of this instrument requires training by the Flow Core faculty. CytExpert software is available for free download only on a Windows system or Virtual PC.

Analysis Software

FlowJo: FlowJo software reads flow cytometry data and facilitates complex data analysis in a graphically intuitive way. FlowJo licenses can be purchased through the Flow Cytometry Core for a fee of $270/year (subject to change).
FCS Express: FCS Express software reads flow cytometry data and facilitates complex data analysis in a graphically intuitive way.
Beckman Coulter Kaluza: Kaluza Software challenges today´s solutions with a new vision of what flow cytometry analysis should be. Kaluza provides visualization tools, speed and an innovative simplicity to the flow community. With features designed to make your life easier in the lab, Kaluza aims to provide a greater understanding of the data generated by modern cytometers. Kaluza employs patent pending technology that allows for real time analysis of high content files allowing you to spend your valuable time on discovery.
CytExpert: Software for the Beckman Coulter CytoFlex data only. CytExpert software is available for a free download. It only runs on a Window system.

There is one first come, first serve basis for a free analysis computer in the core facility that has no sign up times or usage fees. Please notify the staff if the computer has any issues. DO NOT leave any data on this computer. Data will be dumped constantly.

Training

In order to sign up to use machines, you have to be formally trained by the facility. You cannot train others in your lab to use the instrumentation.

Training is available for all Flow Cytometry Analyzers and the FACSAria II Cell Sorter. Trainings are provided by an experienced staff member several times monthly. Also, a "Basics of Flow Cytometry" course is offered 1-2 times per year based on request. In addition, researchers can request Facility run or Facility assisted analysis of samples.
Educational seminars and workshops are provided several times per year for the advancement of the research community.