Flow Cytometry Core
About the Flow Cytometry Core
The Flow Cytometry Core provides state-of-the-art instrumentation and expertise to support studies using high-speed, multiparameter cell analysis and sorting. The Core enables investigators to reliably analyze and isolate purified populations of cells to advance our understanding of diabetes and related metabolic disorders. Training and educational activities of the Core enhance the use of flow cytometry, bringing this technology to new users and stimulating new avenues of investigation.
Core Leadership & Scientific Oversight
The Flow Cytometry Core is led by investigators with expertise in stem cell biology, immunology, and advanced cell analysis techniques, providing scientific direction, operational oversight, and training to support investigators using Core services.
Amy Wagers, PhD, Director, is a stem cell biologist with more than 25 years of experience in multi-parameter flow cytometry and its application to the isolation and analysis of rare cell types. Her research focuses on mechanisms that regulate blood and muscle stem cell function and the development of gene therapy and CRISPR-based approaches in tissues central to metabolic health and diabetes.
Thomas Serwold, PhD, Associate Director, is an immunologist and stem cell biologist with extensive experience in cell sorting technologies. His research focuses on the role of autoimmune T cells in type 1 diabetes.
Angela Wood, Core Manager, is a Senior Research Assistant with extensive experience in flow cytometry operations, instrument troubleshooting, and user training. She supports daily Core operations, including cell sorting and analysis, instrument maintenance, and training programs for new users.
Services
Sorting of cells, nuclei, or similarly sized particles is achieved through instruments that propel a suspension of particles one at a time through a narrow, pressurized fluidics stream, interrogate by sample laser-based excitation of individual particles coupled to emission detection and software-based analysis, and apply a charge to deflect wanted cells or particles into a collection tube. Instruments include:
- SORP BD FACSAria: This instrument is capable of high speed and single-cell sorting on up to 15 fluorescent parameters and is equipped with five lasers (355nm, 488nm, 405nm, 561nm, and 640nm). It is capable of four-way sorting into tubes or single-cell sorting directly into plates or slides. This instrument is equipped with an Aerosol Containment System for the sorting of BSL-2 biohazardous samples.
- BD FACSAria IIu: This self-run instrument is capable of high speed and single-cell sorting on up to 12 fluorescent parameters and is equipped with three lasers (488nm, 405nm, and 640nm). It is capable of four-way sorting into tubes or single-cell sorting directly into plates or slides. This instrument is also equipped with an Aerosol Containment System for the sorting of BSL-2 biohazardous samples.
- BD FACSDiscover S8: Based on input from DRC research base investigators, Joslin is planning to expand its offerings soon with a new instrument that brings on the capabilities for spectral flow sorting and image-based sorting. Spectral deconvolution will enable discrimination of more than 50 different cell features simultaneously and greatly improve the ability to distinguish signal from noise, with enhanced resolution, sensitivity, and compensation that significantly increase the fidelity of flow cytometry and sorting as compared to traditional flow cytometers. In addition, the ability to perform real-time image-enabled sorting, with coincident capture of detailed images of every cell as it passes the detector, will enable studies to quantify changes in cell morphology and spatial organization.
These analyzers are based on FACS principles through a similar process as sorting but without separation of particles into separate collection tubes.
- BD LSR-II: This instrument is equipped with four laser lines (488nm, 405nm, 561nm, and 640nm) and 15 fluorescent detectors. The current configuration allows for two colors off the blue laser, five colors off the yellow-green laser, three colors off the red, and six off of the violet.
- Miltenyi Biotec MACSQuant: The MACSQuant is capable of detecting up to eight fluorescent parameters and is equipped with 488nm, 633nm, and 405nm solid-state lasers. This instrument has the ability to provide absolute cell counts.
- BD LSRFortessa: This instrument is equipped with five laser lines (445nm, 405nm, 488nm, 561nm, and 640nm) and 17 fluorescent detectors, and the HTS (High Throughput Sampler) system which enables fully automated sample acquisition from 96- and 384-well microtiter plates. The current configuration allows for two colors off the 445nm laser, three colors off the blue laser including GFP and YFP detection, five colors off the yellow-green laser, three colors off the red, and four off of the violet.
The CTL ELISPOT reader is designed for scanning and evaluating a wide range of microtiter plate-based bioassays — in particular, ELISPOT assays. It is equipped with both brightfield and fluorescence optics, detecting up to 5 fluorescent channels. This instrument is also well suited for objective reproducible multiwell cell-counting of live cells.
An important function of the Flow Core is to provide expert training, not only for those users who are new to flow cytometry, but also for experienced users who wish to expand their research approaches and push the limits of the technology. It is recommended that potential users consult with Core personnel for fluorochrome combination feasibility and necessary controls prior to designing your analysis experiment.
Data analysis can be accomplished using a number of platforms, and the Core Manager consults with users to guide their analytic approaches. Recognizing that FlowJo software is the analysis package chosen by most users, the Core offers individual annual FlowJo user licenses at a substantially reduced cost.
Chargebacks
Flow Cytometry Core Chargebacks
| Service | Research Base members, academic diabetes or metabolism investigators* | Academic investigators from other fields** | Industry** |
|---|---|---|---|
| Cell Analyzer, self-run | $45/hour | $60/hour | $250/hour |
| Cell Analyzer, operator-assisted | $55/hour | $70/hour | $250/hour |
| Cell Sorter, self-run | $50/hour | $60/hour | $250/hour |
| Cell Sorter, operator-assisted | $60/hour | $70/hour | $250/hour |
| Flow Cytometer use training | $60/hour | $70/hour | $250/hour |
| FlowJo Software | $150/user license | ||
| *Benefit of DRC support accrues to studies relevant to DRC mission. **Investigators from other fields and industry are accommodated as availability permits, but rates do not benefit from DRC support. |
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Core Policies
- The minimum charge for cell sorters is one hour and for analyzers is ½ hour. On all instruments, users are charged for the entirety of their reservation time. This means that if a user is late they will be charged for the time between their scheduled start and their actual arrival. It also means that if a user finishes early, they will still be charged for the remainder of the scheduled time.
- To book time less than 24 hours prior to the start time, users can contact the Joslin Flow Core staff to have them book the time on your behalf.
- All sorting cancellations (self-run or operator-assisted) require 48 hours' notice and all analysis cancellations require 24 hours’ notice and cancellations should be entered into iLab. To cancel time with less than these requirements, contact the Joslin Flow Core staff to cancel the time on your behalf. However, Cancellations made after the deadlines are treated as a no-show. Users will be charged the full usage fees for late cancellations unless someone else schedules for the canceled time. No-shows will incur usage fees for the entire reservation.
Requesting Services
Visit Joslin’s iLab portal to request services.
Contact Us
- Email Angela Wood, Core Manager
- Email Dr. Thomas Serwold, Core Associate Director
- Email Dr. Amy Wagers, Core Director
Remember to Cite the DRC
If research has been supported in full or in part by our Core, users are requested to acknowledge the NIH/NIDDK grant as follows: "Supported by the Flow Cytometry Core of NIH P30 DK036836."
