Our Quantum™ MESF and Quantum™ Simply Cellular® beads are external standards that enable the standardization of fluorescence intensity units irrespective of cytometer and software. Because they are labeled with the same fluorochromes used to label cells, they provide a  synchronous response to the environment. To use, beads are run on the same day and at the same PMT & compensation settings as samples to establish a calibration curve that relates instrument channel values and standardized fluorescence intensity units (MESF or ABC). Cell samples are then read against the curve for determination of expression (i.e. quantitation of the fluor signal from each cell population).

For additional information about working with these products, see our Quantitative Cytometry brochure and our Tips & Troubleshooting PDS or check out our Flow Blog.

 

 

 

Quantum™ MESF

Our Quantum™ MESF kits contain one blank population and a series of either four fluorescent microsphere populations labeled with varying amounts of Alexa Fluor® 488, Alexa Fluor® 647, APC, or R-PE, or five fluorescent microsphere populations labeled with varying amounts of FITC. The assignment of fluorescence intensity in Molecules of Soluble Fluorochrome (MESF) units is performed through direct comparison of fluorescence measurements from solutions of the pure fluorochrome with those from microspheres surface-labeled with the same fluorochrome. See PDS 849 to determine effective FP ratio. A free QuickCal® analysis template is provided with each kit to aid in determining the expression level of cells, and for evaluating instrument linearity and detection threshold. Quantum™ MESF kits are available in three sizes: A (20 tests), B (100 tests), and C (280 tests).

PDS 818, QSC & QMESF Tips and Techniques
PDS 819, QuickCal®
PDS 821, Quantum™ MESF Kits
PDS 849, Determination of Effective F/P Ratio
SDS ST331, (QMESF FITC)
SDS ST332, (QMESF Alexa Fluor® 647)
SDS ST334, (QMESF AF 488, PE & APC)

Catalog NumberDescription
488Quantum™ Alexa Fluor® 488 MESF
555Quantum™ FITC-5 MESF
555pQuantum™ FITC-5 MESF (Premix)
827Quantum™ R-PE MESF
647Quantum™ Alexa Fluor®  647 MESF
823Quantum™ APC MESF

References

Wyant T, Lackey A, Green M. (2008) Validation of a flow cytometry based chemokine internalization assay for use in evaluating the pharmacodynamic response to a receptor agonist. J Transl Med; 6:76.

Quadrini KJ, Hegelund AC, Cortes KE, Xue C, Kennelly SM, Ji H, Högerkorp C-M, Mc Closky TW. (2016) Validation of a flow cytometry-based assay to assess C5aR receptor occupancy on neutrophils and monocytes for use in drug development. Cytometry B Clin Cytom; 90B:177-190. 

Bray RA. (2013) Lymphocyte crossmatching by flow cytometry. Methods Mol Biol; 1034:285-96.

Smith ML, Chyla B, McKeegan E, Tahir SK (2016) Development of a flow cytometric method for quantification of BCL-2 family members in chronic lymphocytic leukemia and correlation with sensitivity to BCL-2 family inhibitors. Cytometry B Clin Cytom

Kay S, Herishanu Y, Pick M, Rogowski, O, Baron S, Naparstek E, Polliack A, Deutsch, V. R. (2006) Quantitative flow cytometry of ZAP-70 levels in chronic lymphocytic leukemia using molecules of equivalent soluble fluorochrome. Cytometry, 70B: 218–226.

Muller L, Mitsuhashi M, Simms P, Gooding WE, Whiteside TL (2016) Tumor-derived exosomes regulate expression of immune function-related genes in human T cell subsets. Sci Rep 6:20254.

Singh H, Figliola MJ, Dawson MJ, Olivares S, Zhang L, et al. (2013) Manufacture of Clinical-Grade CD19-Specific T Cells Stably Expressing Chimeric Antigen Receptor Using Sleeping Beauty System and Artificial Antigen Presenting Cells.PLoS ONE; 8(5):e64138

Homann S, Hofmann C, Gorin AM, Nguyen HCX, Huynh D, Hamid P, ... & Roy SS. (2017) A novel rapid and reproducible flow cytometric method for optimization of transfection efficiency in cells. PloS one, 12(9), e0182941. (Quantum MESF to confirm and quantitate transfection efficiency featuring a fluorescently-labeled vector)

Quantum™ Simply Cellular®

Quantum™ Simply Cellular® (QSC) kits are comprised of five microsphere populations that are ~7-9µm in diameter: one blank and four labeled with increasing amounts of antibody. Each coated population binds a specific number of monoclonal antibodies of the noted species, which is equal to its Antibody Binding Capacity (ABC) value. The investigator labels QSC beads using the same antibodies used to stain cells.  A free QuickCal® analysis template is provided with each kit to aid in determining the expression level of cells, and for evaluating instrument linearity and detection threshold. Quantum™ Simply Cellular® kits are available in three sizes: A (20 tests), B (100 tests), and C (280 tests). For additional information about working with these products, see our product data sheet on Antibody Titration or check out PDS 818, Tips and Troubleshooting.

PDS 814, Quantum™ Simply Cellular®
PDS 814A, QSC/SC Antibody Titration Protocol
PDS 818, QSC & QMESF Tips and Techniques
PDS 819, QuickCal®
SDS ST337, (QSC & SC)

Catalog NumberDescription
815Quantum™ Simply Cellular® anti-Mouse IgG
816Quantum™ Simply Cellular® anti-Human IgG
817Quantum™ Simply Cellular® anti-Rat IgG

References

Bringeland, G. H., Bader, L., Blaser, N., Budzinski, L., Schulz, A. R., Mei, H. E., ... & Gavasso, S. (2019). Optimization of receptor occupancy assays in mass cytometry: Standardization across channels with QSC beads. Cytometry Part A, 95(3), 314-322.

Randlev B, Huang L-C, Watatsu M, Marcus M, Lin A, Shih S-J. (2010) Validation of a quantitative flow cytometer assay for monitoring HER-2/neu expression level in cell-based cancer immunotherapy products. Biologicals; 38(2):249-259.

McBean RS, Wilson B, Liew Y-W, Hyland CA, Flower RL. (2015) Quantitation of Lan antigen in Lan+, Lan+(w) and Lan- phenotypes. Blood Transfus; 13:662-5

Phillips A, Boghaert E, Vaidya K, Mitten M, Norvell S, Falls H, …Reilly A (2016) ABT-414, an Antibody–Drug Conjugate Targeting a Tumor-Selective EGFR Epitope Mol Cancer Ther 15(4)661-669

Tillotson B, Cho YK, Shusta E, (2013) Cells and Cell Lysates: A Direct Approach for Engineering Antibodies Against Membrane Proteins Using Yeast Surface Display Methods 60(1)27–37

Simply Cellular®

Our Simply Cellular® standard consists of a single population of antibody-coated microspheres (~7-9µm in diameter) of known Antibody Binding Capacity (ABC). The population is stained with the user's primary antibody and used in conjunction with a suitable Quantum™ MESF kit for determination of the antibody's effective Fluorophore : Protein (F/P) ratio. This standard may also be used to QC the fluorescence intensity of different antibody lots or clones. Simply Cellular® kits are available in three sizes: A (20 tests), B (100 tests), and C (280 tests). 

PDS 810, Simply Cellular® anti-Mouse IgG, anti-Human IgG, anti-Rat IgG
PDS 814A, QSC/SC Antibody Titration Protocol
PDS 818, QSC & QMESF Tips and Techniques
PDS 849, Determination of Effective F/P Ratio

Catalog NumberDescription
810Simply Cellular® anti-Mouse IgG
812Simply Cellular® anti-Human IgG
813Simply Cellular® anti-Rat IgG

References

Andreev J, Thambi N, Perez Bay AE, Delfino F, Martin J, Kelly M… Thurston G. (2017) Bispecific Antibodies and Antibody–Drug Conjugates (ADCs) Bridging HER2 and Prolactin Receptor Improve Efficacy of HER2 ADCs. Mol Cancer Ther. (16)(4) 681-693 (Quantum MESF PE [quantitation] & Simply Cellular [F:P ratio])

QuickCal®

QuickCal® constructs a calibration curve associating the cytometer's fluorescence channel values to standardized fluorescence intensity units. (MESF or ABC) For instructions regarding QuickCal, please see Product Data Sheet 819, QuickCal® Data Analysis Program or watch our video.