Flow Cytometry Lab

Summary

Our experienced staff leverage the latest cytometry technologies to develop innovative flow cytometry measurement solutions, standards, and reference materials to enable quantitative measurements of biological substances such as cells, extracellular vesicles/exosomes, proteins, viruses, and nucleic acids. We work with the manufacturers of calibration particles and instruments to enable the cytometer standardization and to achieve the detection limit smaller than 20-30 nm in diameter. We have measured 20 nm fluorescently labelled particles and 35 nm unstained particles. The NIST Flow Cytometry Lab is a state-of-the-art facility equipped with four different cytometer platforms.

Description

Current Capabilities

• ERF value assignment measurement service for traceable ERF value assignment to microparticles, Aseptic sorting of cell lines, and primary cells
• Quantitative measurements of cell surface and intracellular biomarkers by estimating antibody bound per cell
• Accurate enumeration of cells with specific phenotypic characteristics and function, e.g., CD34+ stem cells, cytokine producing T cells, and microbes
• Harmonization of flow cytometry data across three different platforms through panel design, antibody titration, instrument calibration, reference cell controls, high dimensional data analysis etc.   

FLOW CYTOMETRY INSTRUMENTS

	Cytoflex flow cytometers from Beckman Coulter Includes up to 6 solid diode lasers, 375 nm, 405 nm, 488 nm, 561 nm, 633 nm and 808 nm and can detect 23 parameters simultaneously on a single sample, 21 color fluorescence data plus forward and side scatter.  Includes Avalanche Photo Diode detector arrays for fluorescence and side scatter light detections and permits the detection of <80nm nano-sized nanoparticles with side-scatter trigger from 405 nm laser.
	Attune NxT from Thermo Fisher Includes 4 solid diode lasers, 405 nm, 488 nm, 561 nm and 633 nm and can detect 16 parameters simultaneously on a single sample, 14 color fluorescence data plus forward and side scatter.  Includes photomultiplier tubes (PMT) for fluorescence and side scatter light detections.
	FACSAria II Sorter from BD Biosciences Includes 3 lasers, 405 nm, 488 nm and 633 nm and can detect 14 parameters simultaneously on a single sample, 12 color fluorescence data plus forward and side scatter.  Includes photomultiplier tubes (PMT) for fluorescence and side scatter light detections. Performs parallel sorting of 4 different live cell populations identified by using 14 parameters.
	Amnis Imagestream MK II Flow Cytometer from Luminex Benchtop multispectral imaging flow cytometer designed for the acquisition of particle imagery with 6 channels. Class 1 laser system with 5 lasers configuration; 405nm (150mW), 488nm (400mW), 561nm (200mW), 633nm (150mw), and 785nm (70mW) Imaging capabilities: 20X, 40X and 60X objective with High Gain functionality for accurate capture of submicron size particles at 60X and Extended Depth of Field (EDF) for advanced spot counting in cells Combines the power of standard microscopy with the statistical significance of large sample data acquisition common to a flow cytometry.
	Aurora Spectral Cytometer from Cytek Includes 5 solid diode lasers, 355 nm 405 nm, 488 nm, 561 nm and 635 nm and can detect up to 40 colors with 67 different parameters and easy detector gain setup. Includes high sensitivity Coarse Wavelength Division Multiplexing (CWDM) semiconductor detector arrays for more efficient spectrum capture Highly sensitive violet SSC, which enables small particle detection and blue laser SSC Vacuum fluidics system allows for flexible sample input