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Flow Cytometry Shared Resource Laboratories

The Flow Cytometry facilities within the College of Medicine are located in the Medical Sciences Building (MSB 2264, 2228) and the Mitchell Cancer Institute (MCI 3085) and both serve as fee-for-service shared resources. The MSB facility is operated by Dr. Domenico Spadafora (over 5 years of experience in flow cytometry) while the MCI facility is operated by Steve McClellan (over 30 years of experience). Dr. Robert Barrington (over 20 years of experience) serves as overall director of both facilities. 

Both facilities house a BD Biosciences FACSAria-SORP cell sorter and a BD FACSCanto II cell analyzer. The FACSAria-SORP cell sorters are capable of 12 fluorescence parameters. The sorter has 4 laser lines, including a 60 mW 350 nm UV line, 100 mW 488 nm, 100 mW 561 nm and 40 mW 640 nm lines. The sorters can be used to isolate cells in multiple formats, including 4-way tube sorting and 96-well plate sorting via the Automated Cell Deposition Unit. The BD FACSCanto II cell analyzers are equipped with 3 lasers (30 mW 405 nm, 20 mW 488 nm, and 17 mW 640 nm) and are capable of 8 fluorescence parameters. 


The MCI Flow Cytometry facility also offers the following:

  • Nexcelom CeligoS microplate-based imaging cytometer: Designed for performing assays directly in multi-well plates (6-384 wells). There are many advantages to this approach, including lower demand for cells (allowing for the use of rare primary cells isolated from patient samples) and reagents; as well as analyzing the cells in their native state without the need to enzymatically remove them from their growth substrate, as is required for traditional flow cytometry. It can image cells in Brightfield and 3 fluorescent channels: blue (377/50 nm excitation; 470/22 nm emission) green (483/32 excitation; 536/40 emission) and orange/red (531/40 excitation; 629/53 emission). Advanced image recognition software provides over 10 specific analysis modes for a wide variety of applications.

  • On-Chip Biotechnologies Microfluidic Cell Sorter (3 laser 6 color): This is world’s first in its class microfluidic cell sorter, developed in Japan and USA MCI was one of the first centers in the US to obtain this new technology. Benefits over traditional sorting include the ability to analyze extremely low starting sample volumes/number of cells. The sorting is much gentler and can accommodate a wide range of particles from exosomes to cell spheres using two different size microfluidic chips. The sorter is aerosol free and placed in a BSL-2 cabinet for safe isolation of cells infected with lentivirus or other pathogens. The 3 excitation lasers (405, 488 & 561 nm) are collinear such that fluorochromes excited by any laser can be detected in any of the following channels: FL1 (445/20 nm), FL2 (543/22 nm), FL3 (591.5/43 nm), FL4 (607/36 nm), FL5 (676/37 nm), and FL6(732/68 nm).

  • On-Chip Biotechnologies Single Cell Plating System (SPiS): As a companion instrument, the SPiS performs single cell or sphere deposition into 96 or 384 well plates in a far more gentle and reliable manner than traditional sorters.

  • XRAD320: X-ray cabinet irradiation systems are designed to deliver precise X-ray dosages to specimens that range in size from cells to small animals.  Both COM SRL locations have nearly identical irradiators (Kimtron IC-320 at MSB; XRAD320 at MCI).  The systems are safe but require assistance and training before independent operation is permitted.  Applications are varied and include production of non-dividing feeder cells, X-ray virus inactivation, production of sterile insects for reducing insect populations, and X-ray sterilization of items that cannot be steam sterilized.

  • ThermoFisher Neon Electroporation System: Designed for primary cells and large size plasmids, very gentle.  Available for use free of charge with appointment.

  • Miltenyi Biotech GentleMACS tissue disassociation system: The industry standard for preparing single cell suspensions from most any tissue with the highest viability.  Available for use free of charge with appointment.


The MSB Flow Cytometry facility also offers the following:

  • ZetaView® TWIN - NTA Nanoparticle Tracking - Video Microscope PMX-220: The 2-laser platform (488 & 640 nm) was installed in the core lab in May 2019. Nanoparticle Tracking Analysis (NTA) captures the Brownian motion of each particle in the video. Based on the different diffusion movements of large and small particles in the surrounding liquid, the hydrodynamic diameter of the particles is determined. Pattern parameters, such as intensity fluctuations, surface geometry and shape of the particles, as well as particle concentration, are documented at each recording and can be used to distinguish sub-populations. In addition, the charge state of the particle surface (zeta potential) can be measured via the movement of the particles in an applied electric field. Depending on the type of sample and the measuring mode, the measuring range is between 15 nm and 5 μm.

  • Agilent Seahorse XFe24 Analyzer: Supported by an S10 award and installed in November 2019, the Seahorse measures the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of live cells in a 24-well plate format. Seahorse XF technology measures the flux of oxygen and protons in the transient microchambers created by specially designed tissue culture microplates in real time. The software calculates the rates of oxygen consumption and extracellular acidification from its measurements of oxygen and proton flux.

  • 10x Genomics Chromium Controller: Installed in February 2020, this instrument uses advanced microfluidics to perform single cell partitioning and barcoding in minutes. The controller combines a single cell with a single gel bead that is coated with a unique oligonucleotide barcode to form a single cell emulsion droplet, which is referred to as a “GEM” (Gel Bead-in-emulsion). Within each GEM, the unique barcoded oligonucleotides are used as primers to generate cDNAs, which can then be traced back to cell of origin. The barcoded amplified fragments from hundreds to thousands of cells are pooled to create short read sequencer compatible libraries. Sequencing of libraries enables analysis of single cells for gene expression, cell surface proteins, immune clonotype, antigen specificity, and chromatin accessibility.

  • Kimtron IC-320: X-ray cabinet irradiation systems are designed to deliver precise X-ray dosages to specimens that range in size from cells to small animals.  Both COM SRL locations have nearly identical irradiators (Kimtron IC-320 at MSB; XRAD320 at MCI).  The systems are safe but require assistance and training before independent operation is permitted.  Applications are varied and include production of non-dividing feeder cells, X-ray virus inactivation, production of sterile insects for reducing insect populations, and X-ray sterilization of items that cannot be steam sterilized.

  • Miltenyi Biotech GentleMACS tissue disassociation system: The industry standard for preparing single cell suspensions from most any tissue with the highest viability.  Available for use free of charge with appointment.

  • Cryostat: A cryostat (Thermo Scientific) is available for sectioning frozen tissues.  Frozen sections can be examined by various histological and immunohistological methods.

  • Finesse 325 Microtome: Formalin-fixed paraffin embedded (FFPE) tissues can be sectioned using the Finesse 325 microtome (Thermo Scientific) located in the MSB facility.  FFPE sections can be examined using various histological and immunohistological approaches.


High-Throughput Compound Screening Core

Located in the Mitchell Cancer Institute and housed within a designated 300 sq. ft. laboratory, this core provides researchers expertise and resources for assay development and high-throughput screening against large compound libraries. The laboratory contains a fully integrated Beckman/Coulter Industrial Robotic Integrated System (IRIS) automated platform for high throughput screening of small molecule libraries fully contained in a HEPA-filtered clean enclosure. The 6-axis Motoman robotic arm and all screening instrumentation are coordinated by the SAMI Workstation EX scheduling software system. Integrated devices include a Molecular Devices ImageXpress Ultra confocal high throughput imager (HCS instrument) capable of rapid, high resolution fluorescent microscopic cellular assays. Automated image acquisition and analysis are controlled by the MetaXpress and Acuity Xpress software packages along with MetaXpress PowerCore Parallel Processing system. An additional Molecular Devices ImageXpress Micro widefield high throughput imager is available for both fluorescent and bright field automated microscopy assays. A BioMek FXp Hybrid Dual Arm automated liquid handler enables transfers of up to 96 samples at once or individual “cherry picking” of active samples and sample titration. A BioTek EL406 microplate washer / dispenser is available for dispensing cells, detection reagents, or processing of immunoassays. The Molecular Devices Paradigm multimode microplate reader enables homogenous endpoint and kinetic detection in absorbance, fluorescent intensity, fluorescent polarization, luminescence, HTRF, and AlphaScreen-based assays. A chemical diversity library of over 50,000 unique chemical entities from Life Chemicals, as well as smaller libraries of known bioactive compounds, and novel focused compound libraries contributed by medicinal chemist collaborators are available for high throughput screening projects.