TECHNOLOGY

Flow cytometry

Enabling high-dimensional, spectral flow cytometry-based cellular phenotyping and precision cell isolation

Technology to drive breakthrough science

The Buck Institute hosts a full-service Flow Cytometry Core, housed in Building 3, Room 324. Our core features two state-of-the-art, 5-laser Cytek spectral cytometry instruments, an Aurora analyzer and an Aurora CS cell sorter, each capable of working with up to 40 simultaneous fluorescence parameters. These platforms enable high-dimensional, high-throughput analysis of single cells based on a host of available fluorescent readouts, allowing for precise isolation and characterization of highly defined and/or rare cell populations.

The parameters we can analyze include expression of surface and intracellular protein markers, reporter genes, and secreted proteins, as well as many other readouts of cellular function such as ion mobilization, protein translation, and mitochondrial function. Our instruments are located in a fully equipped P2* tissue culture suite, allowing processing of time-sensitive samples to be carried out with maximum efficiency. Our sorter also features a biohazard containment system that allows for safe sorting of primary human cells and tissues.

  • Herb Kasler, PhD . Core Director

    Herb Kasler, PhD, is the Director of the Flow Cytometry Core. He got his PhD in immunology from the University of California, Berkeley, working on T cell development in the laboratory of Astar Winoto. Since then he has been working with Eric Verdin, MD, studying gene regulation during T cell development, with a focus on its relationship to autoimmune disease. Flow cytometry has been a cornerstone of all this work, and as a result, Dr. Kasler has more than 20 years of experience using numerous flow-related techniques and platforms. He also received extensive training in the theory, operation, and maintenance of BD cell sorters from Marty Bigos, who served as flow core director for the Gladstone Institutes and later for Stanford University.

    HKasler@buckinstitute.org

  • Ritesh Tiwari, PhD . Flow Core Manager

    Dr. Tiwari completed his masters in biotechnology at Madurai Kamraj University and earned his PhD in immunology from National institute of Immunology, New Delhi, India in 2014 where he worked on the influence of impaired vesicular transport in Beige B cells response. Since then he has worked on multiple projects involving universities and industries helping them educate, design and analyse experimental data involving multi-color flow cytometry. He joined the Verdin lab in September 2019 and his work includes providing support to a diverse array of projects in the biology of aging, both by sorting cells and by helping with immunophenotyping and other types of analyses involving Flow cytometry as well as working on role of chromatin modifications, particularly histone acetylation, in immune aging and autoimmune disease. In his spare time he enjoys going for hike or bike ride with his fellow buck colleagues.

    RTiwari@buckinstitute.org

The core supports the work of Buck scientists in many ways. We can perform full-service deep (>30-parameter) immunophenotyping of both mouse and human specimens, drawing upon our ever-growing collection of antibody reagents and panel design knowledge base. Our high-throughput sterile sorting capability can also support genomic-level screens of expression libraries of cDNAs, shRNAs, or CRISPER guide RNAs, while our high-throughput sampling capability can support screening of libraries of up to 10,000 compounds in a week. Our staff is available for instrument operation, user training, help with sample preparation, and consultation about experimental design and cell isolation strategies. Initial consultations are always free of charge.

External users:

We welcome business from external clients in either academia or industry. Recharge rates will determined on a project-by-project basis.

 

Cytek Aurora spectral flow cytometer: 

  • This state-of-the-art spectral flow instrument features five lasers (355nm, 405nm, 488nm, 561nm, and 640nm) and 64 output channels, enabling highly sensitive detection of virtually any commonly available fluorophore. Violet laser-based scatter detection also enables characterization of particles down to ~300 nm.
  • Panels of up to 40 individual fluorophores can be run, and we routinely run panels of 28-36 fluorophores for immunophenotyping.
  • The integrated 96-well autosampler allows for automated acquisition of up to 200 samples per hour.

Cytek Aurora CS cell sorter: 

  • Five lasers and 64 output channels in addition to real-time spectral unmixing, enabling ultra-precise sorting based on any panel that can be implemented on the analyzer.
  • Sample throughput of up to 60,000,000 cells/hour, depending on nozzle size.
  • Sterile collection of up to 6 simultaneous bulk populations, and sterile sorting into plates up to 384 wells.
  • Biohazard containment up to P2* level.

Other core resources:

  • License dongles for Spectroflo and FlowJo analysis software.
  • A PC data workstation optimized for spectral flow analysis.
  • A large collection of mouse and human antibodies and other flow reagents, available to core users pro-rata.

Please contact us for more detailed information on the Flow Cytometry core capabilities, services and rates.

Herb Kasler
Director, Flow Cytometry Core
hkasler@buckinstitute.org
415-209-2056

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