Macsquant cytometer

Manufactured by Tree Star

Sourced in United States

The MACSQuant cytometer is a flow cytometry instrument designed for high-performance cell analysis and sorting. It is capable of detecting and analyzing multiple parameters of single cells or particles in a fluid sample.

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Lab products found in correlation

Dss1 coolscope slide scanner, by Nikon (2 mentions) Fluoview fv1000 microscope, by Olympus (2 mentions) Tcs sp2 aobs confocal laser scanning microscope, by Leica (2 mentions) Rat anti mouse cd16 cd32 mab, by BD (1 mentions) Fixation buffer, by BioLegend (1 mentions) Permeabilization buffer, by BioLegend (1 mentions) Imagej, by Leica (1 mentions) Confocal software lcs lite, by Leica (1 mentions) Lcs lite software, by Leica (1 mentions) Cd161 pe, by Miltenyi Biotec (1 mentions) Ix permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Cd3 fitc, by BioLegend (1 mentions) Cd19 bv421, by BD (1 mentions) Cd56 apc, by BioLegend (1 mentions) Cd38 percp cy5, by BioLegend (1 mentions) Cd8 percp cy5, by BioLegend (1 mentions) Cd4 viogreen, by Miltenyi Biotec (1 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (1 mentions)

5 protocols using macsquant cytometer

Isolation and Characterization of Tumor-Associated Macrophages

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Tumor tissues were harvested, minced, and then incubated with digestion buffer (RPMI supplemented with 3 mg/ml Dispase II, 1 mg/ml Collagenase I, Clostridium Histolyticum) for 30 min at 37°C. Digestion mixtures were filtered through 0.7 µm nylon strainers (BD Falcon), washed twice with cold PBS, and then incubated for 10 min at 4°C with rat anti-mouse CD16/CD32 mAb (BD Biosciences) to block nonspecific binding. Cells were then stained with anti-CD206 antibody conjugated with Alexa488 (BioLegend) and anti-F4/80 antibody conjugated with APC (BioLegend), followed by washing with PBS and fixation with 1% paraformaldehyde. Data acquisition and analysis were performed on a MACSquant cytometer using FlowJo software (Treestar Inc.).

Fang J., Xiao L., Joo K.I., Liu Y., Zhang C., Liu S., Conti P.S., Li Z, & Wang P. (2015). A Potent Immunotoxin Targeting Fibroblast Activation Protein for Treatment of Breast Cancer in Mice. International journal of cancer. Journal international du cancer, 138(4), 1013-1023.

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Multiparameter Flow Cytometry Protocol

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For surface staining, cells were suspended in FACS buffer (1% BSA in PBS) and incubated with purified antibodies for 30 min at 4°C, then washed 3x in FACS buffer and fixed in 0.5% paraformaldehyde. For intracellular staining, cells were stained for surface markers and fixed with a 1:4 dilution of Fixation Buffer (Biolegend) in PBS for 20 minutes; washed twice with a 1:10 dilution of Permeabilization Buffer (Biolegend) in PBS (Perm Buffer); incubated in Perm Buffer for 20 minutes; stained for CXCL9 or IFNγ in Perm Buffer for 30 min; and washed twice with FACS buffer. All data were acquired with a Miltenyi MacsQuant cytometer and analyzed with FlowJo (TreeStar) software.

Clancy-Thompson E., Perekslis T.J., Croteau W., Alexander M.P., Chabanet T.B., Turk M.J., Huang Y.H, & Mullins D.W. (2015). Melanoma induces, and adenosine suppresses, CXCR3-cognate chemokine production and T-cell infiltration of lungs bearing metastatic-like disease. Cancer immunology research, 3(8), 956-967.

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Multimodal Cytometry and Microscopy Analysis

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FACS analysis was performed on Miltenyi Biotec MACSQuant cytometer and analyzed with FlowJo version 9.6.2 software (Tree Star, Ashland, OR). Graphs and statistical analysis were performed using GraphPad Prism version 6.0a (GraphPad Software, San Diego, CA).
For analysis of immunohistochemical staining, images were acquired on a DSS1 Coolscope slide scanner (Nikon, Kingston upon Thames, U.K.).
For immunofluorescent microscopy, images were acquired on an Olympus Fluoview FV1000 microscope (Olympus, Southend-on-Sea, U.K.) and analyzed using Fiji (ImageJ, National Institutes of Health, Bethesda, MD). The Leica TCS-SP2 AOBS confocal laser-scanning microscope was used for the CD161 immunofluorescence, and the images were captured and processed with Leica confocal software (LCS Lite) and ImageJ.

Llibre A., López-Macías C., Marafioti T., Mehta H., Partridge A., Kanzig C., Rivellese F., Galson J.D., Walker L.J., Milne P., Phillips R.E., Kelly D.F., Freeman G.J., El Shikh M.E., Klenerman P, & Willberg C.B. (2016). LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation. The Journal of Immunology Author Choice, 196(5), 2085-2094.

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Multimodal Cytometry and Microscopy Protocol

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FACS analysis was performed on Milteny Biotec MACSQuant cytometer and analysed with FlowJo Version 9.6.2 software (TreeStar, USA). Graphs and statistical analysis were performed using GraphPad Prism Version 6.0a (GraphPad Software, USA).
For analysis of immunohistochemical staining, images were acquired on a DSS1 Coolscope Slide Scanner (Nikon, UK).
For immunofluorescent microscopy, images were acquired on an Olympus Fluoview FV1000 microscope (Olympus, UK) and analysed using Fiji (ImageJ, National Institute of Health, USA). The Leica TCS-SP2 AOBS confocal laser-scanning microscope was used for the CD161 immunofluorescence and the images were captured and processed with Leica Confocal, LCS Lite software, and Image-J.

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Multicolor Flow Cytometry Immunophenotyping

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For external staining, cells from cell lines or PBMCs in PBS were incubated with anti-surface antibodies at room temperature (RT) for 20 min. Live/dead staining was performed using LIVE/DEAD® Fixable Near-IR Dead Cell Stain Kit(Invitrogen), at 633 or 635 nm excitation.
For internal staining, cells were fixed with 2% formaldehyde (Sigma Aldrich) in PBS for 10 min and permeabilized with IX permeabilization buffer (eBioscience) in water.
The following antibodies were used: CD3-FITC (BioLegend, Catalog No. 300406, clone UCHT1, Mouse IgG1, k), CD8-PerCP-Cy5.5 (BioLegend, Catalog No. 344710, clone SK1, Mouse IgG1, k), CD38-PerCP-Cy5.5 (BioLegend, Catalog No. 303522, clone HIT2, Mouse IgG1, k), CD56-APC (Biolegend, Catalog No. 318310, clone HCD56, Mouse IgG1, k); CD19-BV421 (BD Bioscience, Catalog No. 562441, clone HIB19, mouse IgG1, k); CD4-VioGreen (Miltenyi Biotec, Catalog No. 130-106-712, clone M-T466, Mouse IgG1, k), CD161-PE (Miltenyi Biotec, Catalog No. 130-092-677, clone 191B8, Mouse IgG2a), IgG2A isotype control (R&D Systems, Catalog No. MAB003, mouse); and 2H7 mAb. When non-conjugated primary antibodies were used, a secondary rat anti-mouse IgG2A-PE (R&D Systems, Catalog No. F0129, clone 344701, IgG1) was used.
FACS analysis was performed on Miltenyi Biotec MACSQuant cytometer and analyzed with FlowJo Version 9.6.2 software (TreeStar).

Llibre A., Garner L., Partridge A., Freeman G.J., Klenerman P, & Willberg C.B. (2016). Expression of lectin-like transcript-1 in human tissues. F1000Research, 5, 2929.

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