Quartz cuvette gel coupled to 1.2 numerical aperture (NA) collection lens, 200 x 200 μm Instant “on/off” reduces usage and/or aging by 10x only turns on when acquiring samples reports hours of usageįlat-top laser specified at the flow cellĬoefficient of variation (CV) <3% over the width of the flat-top laser No warm-up delay fiber isn’t affected by “on/off” Up to 14 color channels with wavelength-tuned photomultiplier tubes (PMTs) user-changeable, keyed filtersįixed alignment with prealigned welded fiber no user maintenance required Optimized excitation for minimized stray laser-line noise and losses to reflectionġ0 x 50 μm flat-top laser providing robust alignment ![]() * Amount of measured usable laser power after light has gone through the beam optics and shaping filters. (I) Finally, NK cells lack B cell and T cell markers (CD19 –CD3 –) and express CD56. (H, K) CD62L identifies naive (T N) CD4 and CD8 T cells, while HLA-DR is expressed by activated T cells (T A). (G) T cells can be further subdivided into CD4 + (T helper cells) and CD8 + (cytotoxic T cells) subpopulations, while (J) regulatory T cells express CD4 and CD25. (E) B cells can be further characterized by HLA-DR and CD45RA expression. (F) Within the lymphocyte gate, immune cells can be subdivided based on their expression of CD3 (T cells), CD19 (B cells), or neither (NK cells). (D) Monocytes are found above lymphocytes on the scatter plot and express both CD14 and CD33. (C) Lymphocytes and monocytes were identified based on forward and side scatter profiles. (B) CD45 + gating was used to select the leukocyte population from the lysed whole blood. (A) Dead cells were excluded from the analysis by gating on live (propidium iodide –) cells in a dot plot. The concentration statistic representative of events/μL of any given population and percent positive are examined in depth.Gating strategy for 13-color immunophenotyping analysis of stained human whole blood using a stain/lyse protocol. Human whole blood cells were stained as described in the application note and acquired and analyzed on the Attune NxT Flow Cytometer. ![]() The labeled cells were fixed, then added in equal amounts to each well of two 96-well plates, and acquired on a 4-laser Attune NxT Flow Cytometer with Autosampler. In brief, lysed human whole blood was labeled with fluorophore-conjugated monoclonal antibodies for CD45, CD3, CD4, and CD8 targets. In this application note, consistency across 96-well plates was examined. These recent developments expand the role of flow cytometry in drug discovery and life science research and are a key factor in understanding cellular and molecular networks. Given time savings, high content, and assay robustness, high-throughput flow cytometry provides the capacity needed as part of an effective and practical systems biology approach. Samples are introduced to the Attune NxT Flow Cytometer with syringes, producing accurate measurements of the volumes of acquired samples and thus accurate calculation of cell concentrations. The Attune NxT Flow Cytometer uses a unique volumetric sample and sheath fluid delivery system. In addition, up to 16 different parameters per cell can be measured on the Attune NxT Flow Cytometer, and thousands to millions of cells can be analyzed in minutes at up to 35,000 cells/sec and 1 mL/min sample input. ![]() This can be performed using the Invitrogen™ Attune™ NxT Flow Cytometer with Attune™ NxT Autosampler, where 96- and 384-well plates can be acquired and analyzed. High-throughput flow cytometry is a particularly useful approach for screening compounds in cell models where multiple readouts are desired. The screening of molecules for effects in living cell populations has become an integral step in virtually every drug discovery program. The emerging field of high-throughput (HT) flow cytometry is extending the capabilities of cell-based screening technologies into the profiling of compound libraries.
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