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Sylvester Flow Cytometry Shared Resource

University of Miami

Contact Info:

Eric Wieder, PhD

Rosenstiel Medical Science Building

1600 NW 10th Avenue. Suite 7147

Miami, FL 33136

https://www.sylvester.org/fcsr

Grants and Identifiers:

RRID: RRID:SCR_022501

Other Citation Identifiers

P30-CA240139

FL DOH 8BC09

Instrumentation:

BD FACSAria IIu High Speed Cell Sorter

Aria has four lasers, 405, 488, 561 and 633 nm, and can detect light in sixteen photomultiplier tubes. Its primary function is to examine complex populations of cells and yield pure populations of cells. The Aria can sort lymphocytes at a rate of approximately 15,000 events per second. For larger cells, the rate is much slower in order to optimize the passage of cells through the instrument. Cells from single populations can be sorted into 96-well plates. Alternatively, the Aria can sort into two 15-ml tubes or four 5-ml tubes. The Aria can purify samples that are simply positive and negative for a single fluorophore or as complex as nine-color samples with intricate gating schemes. The BD FACSAria IIu cell sorter is a high speed benchtop digital flow cytometer equipped with a sensitive, fixed-alignment cuvette flow cell. It has three spatially separated lasers - 405 nm, 488 nm and 633 nm. Its primary function is to analyze complex populations of cells and yield pure populations of cells meeting defined criteria. The sorter can purify samples that are simply positive and negative for a single fluorophore or as complex as multi-color samples with complex gating strategies. Sorting can be performed into two or four tubes with speeds up to 25,000 events/second. The system sorts by incorporating cells from the sample tube into a stream of sterile PBS. The stream is interrogated by the lasers at the flow cell and the system electronics keeps track of each cell as they pass through the laser and determines specific cells that meet the sort criteria. A transducer vibrates the stream and induces droplet formation, with cells in the stream being incorporated into the droplets. If a cell meets the sorting criteria and is in the last drop before the break off, the instrument will charge that drop. The charged droplet is then deflected into the proper collection tube by the charge plates. Different cell types can be sorted with the use of 70 ?m, 85 ?m and 100 ?m nozzles. If you need a yellow-green (561 nm) laser, check the BD FACSAria III instead. [Product Link]

RRID:SCR_019598

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BD LSRFortessa Fortessa Flow Cytometer

BD LSRFortessa cell analyzer offers the ultimate in choice for flow cytometry, providing power, performance, and consistency. Designed to be and expandable, the BD LSRFortessa has the flexibility to support the expanding needs of multicolor flow cytometry assays. The BD LSRFortessa system can be configured with up to 7 lasers*,blue, red, violet, UV and yellow-green. The instrument can accommodate the detection of up to 18 colors simultaneously with a defined set of optical filters that meet or exceed the majority of today's assay requirements. BD FACSDiva software controls the efficient setup, acquisition, and analysis of flow cytometry data from the BD LSRFortessa workstation. The software is common across BD FACS instrument families, including the BD FACSCanto cell analyzer and BD FACSAria cell sorter systems. [Product Link]

RRID:SCR_019601

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BD Scientific FACSCanto™ Flow Cytometer

The BD FACSCanto™ is a flow cytometry system that performs reliable performance, accuracy, and ease-of-use for clinical diagnostic services. The BD FACSCanto system has 10-color capability, optical enhancements, and a 4-3-3 configuration deliver high sensitivity and resolution for accurate results. [Product Link]

RRID:SCR_018055

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Fluidigm CyTOF Helios Mass Cytometer

Mass cytometer that enables deep profiling of translational and clinical research samples across a range of cell surface and intracellular markers. [Product Link]

RRID:SCR_019916

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BD FACSymphony A5 SE

Flow Cytometer

Beckman Coulter CytoFLEX SRT

cell sorter

Fluidigm Hyperion

Imaging Mass Cytometer

Other Equipment

BD FACSAria Fusion Cell Sorter Beckman Coulter CytoFLEX SRT Cell Sorter BD FACSymphony A5 SE Flow Cytometer ALS CelCellector Robotic cell/colony picker

Services Provided:

Cell Sorting

CyTOF

Data Analysis

Data Analysis And Interpretation

FACS Cell Sorting

Flow Cytometric Analysis

Flow Cytometry Data Analysis

Imaging Mass Cytomety

Mass Cytometry

Reagents

Relevant Publications:

1.) Garikapati K, Young IC, Hong S, Rai P, Jain C, Briegel KJ (2024 Mar). Blocking LBH expression causes replication stress and sensitizes triple-negative breast cancer cells to ATR inhibitor treatment. Oncogene, 43(12), 851-865. doi: 10.1038/s41388-024-02951-3. PMID: 38297083.


2.) Kurtenbach S, Sanchez MI, Kuznetsoff J, Rodriguez DA, Weich N, Dollar JJ, Cruz A, Kurtenbach S, Field MG, Durante MA, Decatur C, Sorouri M, Lai F, Yenisehirli G, Fang B, Shiekhattar R, Pelaez D, Correa ZM, Verdun RE, Harbour JW (2024 Feb). PRAME induces genomic instability in uveal melanoma. Oncogene, 43(8), 555-565. . PMID: 38030788.


Description:

The Flow Cytometry Shared Resource (FCSR) at Sylvester is an ISAC-recognized Shared Resource Laboratory for excellence, and provides researchers with sophisticated methods for the analysis and preparative sorting of both normal and tumor cells. The facility helps researchers to measure parameters including apoptosis, gene expression, drug metabolism, immune responses and pathways of cellular activation, both normal and tumor-related. The facility also provides access to high parameter cytometry instrumentation including spectral cytometry, mass cytometry (CyTOF) and imaging mass cytometry,