Flow Cytometry Core
The Department of Pharmacology and Toxicology Flow Cytometry Core:
The mission of the Department of Pharmacology and Toxicology Flow Cytometry Core is to provide investigators access to cell sorting and analytical flow cytometry services, as well as training and experimental consultation.
BD FACS Aria IIu
The BD FACS Aria IIu is the original model FACS Aria-I cell sorter that has been upgraded with a FACS Aria II flow cell. It is equipped with 3 lasers: a blue 488nm laser (6 detectors + FSC/SSC), a red 633nm laser (3 detectors), and a violet 405nm laser (4 detectors). The optical configuration allows multi-parameter detection of up to 13 fluorescent parameters and 2 light scatter parameters. The FACS Aria-IIu is capable of sorting through over 50 million cells per hour (dependent on cell type), with up to four populations simultaneously. The populations of interest can be sorted into 0.5 mL/1 mL Eppendorf tubes, 5 mL tubes, or 15 mL conical tubes. Populations can also be sorted directly into multi-well plates, including: 12-well, 24-well, 48-well, 96-well.
BD Accuri C6
The BD Accuri C6 is a compact flow cytometer that uses a low-pressure pumping system to drive the fluidics allowing for the derivation of sample volume and calculation of absolute counts or sample concentration per microliter. The instrument is capable of running up to 10,000 events per second at sample concentrations of over 5 x 106 cells/mL. It is equipped with 2 lasers: a blue 488nm (3 detectors + FSC/SSC; FL1 [530±15 nm], FL2 [585±20 nm], FL3 [>670 nm]) and a red 633nm (1 detector; FL-4 [675±12.5 nm]). The optical configuration allows multi-parameter detection of up to 4 fluorescent parameters and 2 light scatter parameters. Optical filters allow for detection of commonly used fluorescent markers, including, but not limited to:
FL1- FITC, AF488, GFP, CFSE, YFP
FL2- PE, PI
FL3- PerCP, PerCP-Cy5.5, PE-Cy7, RFP, mCherry
FL4- APC, AF647, APC-Cy7, APC-H7
The Luminex 200 is a diagnostic instrument that enables bead-based multiplex analysis of up to 100 analytes simultaneously in a single low-volume (≥ 2 µL) sample. This system utilizes fluorescently labeled microbeads that are conjugated with a specific reagent (eg, antibody, antigen, enzyme substrate, etc.) enable capture and detection of analytes of interest. The Luminex 200 is equipped with 2 lasers: a 635nm classification laser for excitation of fluorescently-labeled microbeads and a 532nm reporter laser for excitation of the analyte signal. The Luminex 200 has a dynamic range typically ≥ 3.5 logs.
Commercially available and custom multiplex kits can be analyzed on the Luminex 200 to quantify protein expression (ie, cytokines, growth factors, phosphoproteins, etc.) in a variety of species (ie, human, mouse, rat, dog, etc.) and samples types (ie, cell culture media, plasma, serum, BALF, etc.). In addition to immunoassays, the platform can be used for enzyme assays, genotyping, gene expression analysis, or assessment of protein-protein interactions.
A handheld magnetic plate washer, microtiter plate shaker, bath sonicator, and vortexer are available in the core to enable multiplex assay performance.
Department of Pharmacology & Toxicology Flow Cytometry Core Billing:
- User reserves instrument time on iLabs providing relevant information in the questionnaire and account number(s) to be charged.
- Once the user finishes running samples and appropriately cleans the instrument, run time is logged manually in the User Log.
- At the end of the month, the amount of time reserved on iLabs and the amount of time logged for each reservation will be reviewed. The investigator will be billed for whichever amount of time is greater for each use. The total amount of time used each month is then listed by user and sent to the department for bills at the end of the month.
Department of Pharmacology & Toxicology Flow Cytometry Core Policies:
- The BD FACS Aria IIu and Luminex instruments use can be scheduled between normal business hours: Monday-Friday from 8am-5pm.
- Scheduling for assisted cell sorting or Luminex use must be booked at least 48 hours in advance.
- For sorting: A charge for the full scheduled usage time will be incurred if reservations are cancelled less than 48 hours in advance of scheduled instrument use.
- BD FACS Aria IIu: An initial consultation ($64.69/hr) is required prior to a preliminary sort in order to discuss experimental design, required samples/controls, and best accommodate the investigator’s and operator’s needs.
- Luminex 200: An initial one-time training session fee ($97.05) is required prior to the 1st use of the Luminex system.
- For analytical use of the BD FACS Aria IIu: A charge for the full scheduled usage time will be incurred if reservations are cancelled less than 24 hours in advance of scheduled instrument use.
- No charges will be applied for cancelled time, regardless of when it was cancelled, if another user schedules that time.
- If you are more than 20 minutes late and have not provided advanced notification, the core reserves the right to cancel your sort and bill you for the reserved time.
- For unassisted use of instrumentation, shifting your time on day of use is acceptable at no charge as long as you will not overlap another user’s scheduled time. Please update your scheduled time on iLabs prior to using instrumentation.
- For assisted use of instrumentation, including sorts, any shift in time of instrumentation use will result in billing according to the scheduled time, plus any additional time incurred.
- In the event the instrument you are scheduled to use is non-operational, the core will notify you as soon as possible.
- If your actual run time exceeds your scheduled time and runs into another user’s time, the subsequent user has the right to bump you to begin their run at the scheduled time.
- Follow all posted cleaning guidelines that are available in associated instrument SOPs or posted by the instrument.
Instrument use can be booked on iLabs
The Pharmacology and Toxicology Flow Cytometry Core Facility is located in the Life Science Building, Room B431.
Please contact Matt Bernard to setup an initial consultation: email@example.com