Flow Cytometry
Flow Cytometry
The Flow Cytometry Core maintains a suite of instruments and software applications for the analysis and sorting of cells and small particles based on the detection of fluorescently labeled markers. Expert cytometry staff are available for training users in the operation of instruments and analytic software, to assist with cytometry-focused experimental design, and perform sample analysis and cell sorting on critical experimental projects.
All new users of the Core are required to undergo training conducted by Core staff prior to independent use of instrumentation. Training can be initiated through the iLabs portal by filling out a “Flow Cytometry Training Request” under the “Request Services” tab, or by selecting “Initiate Training” next to the instrument of interest. After training is complete, users are invited to freely reserve time on instruments using the calendars in the iLabs portal under “Schedule Equipment” (which can be directly accessed following the links provided below).Analyzer instruments (such as the Accuri, Celesta, LSRII, Symphony, and ImageStream) and the Melody sorter are reserved by users on a first-come, first-served basis and are available 24/7. Staff-assisted cell sorting appointments on the Aria cell sorter are also be booked by users after completing a sorting consultation prior to a first appointment (submit a “Flow Cytometry Aria Cell Sorting Consultation” request). Aria sorting appointments must be booked or canceled at least 12 hours prior to the intended time of use. If last-minute bookings or cancellations are needed, please reach out to Core staff directly to inquire.Users are highly encouraged to consult Core staff for experimental design recommendations and sample preparation prior to utilizing instrumentation. For critical experiments or when additional assistance is needed, we strongly suggest planning experimental procedures to fall within the staffed time window of the core (Monday-Friday, 9:00am-5:00pm) and to verify with Core staff of their availability. Please contact Core staff well in advance (1-2 weeks minimally) about your needs or to inquire about special accommodations.
Instrumentation and Services
Aria Cell Sorter
THIS INSTRUMENT IS SPECIALIST-OPERATED ONLY. Droplet-based cell sorter from BD Biosciences. This device has four lasers (405, 488, 561 and 633 nm) and can analyze up to nine fluorescent parameters through conventional cytometry methods. Users may book appointments freely, as available on the schedule. Flow Core staff review all appointment requests and promptly send confirmation of approval. If booking for new sample types or design panels, please reach out to Flow Core staff via email or submit an Aria Cell Sorting Consultation request under the “Services” tab.
Melody Cell Sorter
Droplet-based cell sorter from BD Biosciences. This device has three lasers (405, 488 and 633 nm) and can analyze up to nine fluorescent parameters through conventional cytometry methods. Samples for sorting must be prepared in 5 mL round-bottom (FACS) tubes and filtered through 30 or 70 um strainers just prior to loading on the device (see detailed sample prep guide for more information). Samples can be sorted as 1-4 unique populations into 5 mL round-bottom (FACS) tubes (1-4), 1.5 mL microcentrifuge tubes (1-4), 15 mL conical tubes (1-2) or various well-plate formats (1).
Accuri
Simple, conventional flow cytometer from BD Biosciences. This device as two lasers (488 and 640 nm) and can detect up to four fluorescent parameters, along with FSC and SSC. Samples can be run manually or automatically in 5 mL round-bottom (FACS) tubes or various well-plate formats.
Celesta
Basic, conventional flow cytometer from BD Biosciences. This device has four lasers (405, 488, 561 and 633 nm) and can detect up to 14 fluorescent parameters, along with FSC and SSC. In comparison to the LSRII, the Celesta can detect numerous Brilliant Violet dyes. Samples can be analyzed manually in 5 mL round-bottom (FACS) tubes or automatically in various well-plate formats utilizing the attached HTS device.
LSRII
Basic, conventional flow cytometer from BD Biosciences. This device has four lasers (405, 488, 561 and 633 nm) and can detect up to 11 fluorescent parameters, along with FSC and SSC. Samples can be analyzed manually in 5 mL round-bottom (FACS) tubes or automatically in various well-plate formats utilizing the accompanying HTS device.
Symphony A5 SE
Spectrally enabled flow cytometer from BD Biosciences. This device has five lasers (355, 405, 488, 561 and 637 nm) and 50 fluorescent detectors. In comparison to strictly conventional flow cytometers (e.g., Celesta), the Symphony can potentially resolve similar fluorochromes from one another (e.g., APC vs. AF647). Samples can be analyzed manually in 5 mL round-bottom (FACS) tubes or automatically in various well-plate formats utilizing the attached HTS device.
ImageStreamX
Imaging flow cytometer from Cytek (formally Amnis). This device has three lasers (405, 488 and 642 nm) and six detector channels. The detector channels can capture up to six fluorescent markers, but it is recommended to designate at least one channel to brightfield imaging and the sixth channel to SSC analysis. Samples are analyzed in 1.5 mL microcentrifuge tubes.
Flow Cytometry Analysis Platforms
The Core provides users with free access to computer workstations containing the latest versions of FlowJo and Ideas software for general flow cytometry and imaging cytometry analysis, respectively. These workstations can be reserved via the equipment calendars under “Schedule Equipment” or used in a walk-up fashion. Training on cytometry software and appointments for assisted sample analysis can be initiated by filling out a “Flow Cytometry Training Request” or “Flow Cytometry Assisted-Use Request” under the “Request Services” tab in the iLabs portal. To inquire about personal FlowJo license access for yourself or your lab, please contact Core staff directly.