Acoustic-assisted hydrodynamic focusing technology and advanced fluidics are designed to minimize clogging and effectively handle a broad range of cell types. This allows for a higher degree of data, detail, and throughput that enables processing of a large range of sample types, including large clumpy cells, samples with a low concentration of cells, and precious samples, more quickly and accurately than ever before with no loss in data quality.
Today fluorescent proteins are widely used in the investigation of gene expression as well as protein localization, translocation, and trafficking within live cells. More advanced techniques include assessment of protein–protein interactions and spatial relationships of proteins in live cells using fluorescence resonance energy transfer (FRET) techniques and fluorescence lifetime imaging microscopy (FLIM).
The simultaneous detection of multiple fluorescent proteins in the same cell has traditionally been more difficult than the detection of multiple fluorophore-labeled antibodies. This is in part because fluorescent proteins have a different, broader emission spectrum than the traditional cell dyes and fluorophores used in the labeling of antibodies.
The Attune NxT Flow Cytometer was developed with fluorescent protein analysis in mind; it enables easy and accurate analysis of multiple fluorescent proteins and fluorescently labeled antibodies (separately or in combination), with configurations allowing up to 4 lasers and 16 detection channels.
The Attune NxT flow cytometer can be used quickly and effectively analyze the editing efficiency in CRISPR edited cells. Analysis by flow cytometry offers several advantages as compared to other methods of editing efficiency analysis:
Analysis of CRISPR edited cells using the Attune™ NxT provides accurate, and rapid quantification of editing efficiency, and is particularly beneficial when multiplexing multiple CRISPR gRNAs. Single cell analysis, functional knockout efficacy, quick actionable data, and minimal samples processing time are a few of the benefits of using flow cytometry for analysis of gene editing for flow cytometry.
Flow cytometry a high-throughput, rapid, and accurate method for quantify functional protein knockdown in CRISPR-edited cell populations. Flow cytometry is particularly beneficial when analyzing cell populations edited with multiple gRNAs, as it provides the ability to ascertain single cell protein knockdown efficiency at multiple loci, without the need for clonal isolation.
The workflow is streamlined, requires minimal reagents and hands-on time and provides rapid and accurate results.
With the Attune NxT Flow Cytometer you can achieve a reliable measure of accuracy for detection of cell populations comprising less than 1% of the total cells by easily running large sample volumes in a fraction of the time without the need to concentrate your sample (below).
Flow cytometry is the method of choice for identifying cells within complex populations, as it allows for multi parameter analysis of thousands to millions of cells in a short time. Strong signal separation in the Attune NxT Flow Cytometer shows excellent separation of cell populations into subsets for immunophenotyping. A wide range of reagent choices as well as the system’s automated compensation module, 4 spatially separated lasers and 14 color choices help simplify multi-color panel design.
The data below describes 13-color immunophenotyping analysis of stained human whole blood using a stain/lyse protocol on the Attune NxT Flow Cytometer. Lymphocyte, monocyte and granulocyte populations were distinguished with forward scatter (FSC) and side scatter (SSC); and monocyte, T cell, B cell and NK populations were identified using fluorescent antibodies against surface antigens specific for the different immunological populations.
On traditional flow cytometers, very dilute samples can take a long time to acquire due to slower flow rates. The Attune NxT Flow Cytometer can run very dilute samples quickly (below).
Providing a simple, fast, accurate, and reliable methodology, flow cytometry has become the method of choice to determine C-values in plant homogenates, and the use of flow cytometry in plant biology has increased rapidly. The Attune NxT Flow Cytometer is well suited for DNA content evaluation. Any of the standard configurations may be used, including the most affordable single-laser system.
Resting platelets, are the smallest cellular component of peripheral blood. Upon activation, platelets undergo rapid changes in cell surface receptor expression that lead to altered adhesive properties and changes in morphology that promote the formation of a platelet plug at the site of vascular disruption. These properties can make the interrogation of platelets by flow cytometry challenging, especially in the context of light scatter detection.
The Attune NxT Flow Cytometer, along with the Attune NxT No-Wash, No-Lyse Filter Kit for violet laser SSC detection, offers a robust assay for detecting platelets in whole blood without sample manipulation. The system’s acoustic focusing technology empowers research with unmatched speed (up to 10 times faster than the traditional cytometers), thereby greatly reducing the assay time.
The ability to direct human pluripotent stem cells (hPSCs) toward differentiated cell phenotypes offers tremendous potential for personalized and regenerative medicine. The Attune NxT Flow Cytometer is ideally suited for use with fragile and large cell types like stem cells and cardiomyocytes (below). Engineered to actively resist clogging, a syringe-driven system and larger flow cell help prevent the loss of precious sample and is drastically less susceptible to clogs.
Memory antigen specific CD4 T cells are quite rare in the circulating blood, with frequency ranging from 1 in 100 to less than 1 in 100,000 depending on the antigen and normal range variation. Flow cytometry is an effective technology to monitor and identify rare cells among a mixed population of different cells types. Not only is it capable of rapidly identifying unique cell types, but it can also be used to analyze many other phenotypic features at the single-cell level, making it a valuable tool for understanding the immune system.
In this study, a viability dye (Invitrogen LIVE/DEAD Fixable Near-IR Dead Cell Stain) and 7 antibodies, including CD137 and CD69, were used as a backbone panel to identify antigen specific CD4 T cells using the Attune NxT Flow Cytometer, 4-laser configuration.
Interest in regulatory T cells has been accelerated by evidence from experimental mouse and human models demonstrating that the immunosuppressive potential of these cells can be utilized in research associated with autoimmunity, infectious agents, and cancer.
Strong signal separation in the Attune NxT Flow Cytometer shows excellent separation of cell populations into subsets for immunophenotyping. This 3-color immunophenotyping analysis of stained mouse splenocytes using the Foxp3 Transcription Factor Staining Buffer Kit shows excellent cell population resolution for mouse regulatory T cells consisting of both surface and intracellular markers.