Our Quantum™ MESF and Quantum™ Simply Cellular® microspheres are external standards that enable the standardization of fluorescence intensity units irrespective of instrument and software. Moreover, they are labeled with the actual fluorochromes used to label cells, for synchronous response to the environment. The beads are run on the same day and at the same settings as samples to establish a calibration curve relating instrument channel values and standardized fluorescence intensity units. Unknowns may then be read against the curve for determination of expression (i.e. quantitation of the signal from each cell population). For additional information about working with these products, see our Quantitative Cytomtery brochure or check out PDS 818, Tips and Troubleshooting.

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, Cy™5, PE-Cy™5, 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. 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).

Catalog CodeDescriptionProduct Data SheetSafety Data Sheet
488Quantum™ Alexa Fluor® 488 MESFPDS 821.pdf & PDS 818SDS ST334.pdf
555Quantum™ FITC-5 MESFPDS 821.pdf & PDS 818SDS ST331.pdf
555pQuantum™ FITC-5 MESF (Premix)PDS 821.pdf & PDS 818SDS ST331.pdf
827Quantum™ R-PE MESFPDS 821.pdf & PDS 818SDS ST334.pdf
828Quantum™ PE-Cy™5 MESFPDS 821.pdf & PDS 818SDS ST334.pdf
822Quantum™ Cy™5 MESFPDS 821.pdf & PDS 818SDS ST334.pdf
647Quantum™ Alexa Fluor®  647 MESFPDS 821.pdf & PDS 818SDS ST332.pdf
823Quantum™ APC MESFPDS 821.pdf & PDS 818SDS ST334.pdf

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 doi: 10.1002/cyto.b.21383

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.

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.

Catalog CodeDescriptionProduct Data SheetSafety Data Sheet
815Quantum™ Simply Cellular® anti-Mouse IgGPDS 814.pdf & PDS 818SDS ST337.pdf
816Quantum™ Simply Cellular® anti-Human IgGPDS 814.pdf & PDS 818SDS ST337.pdf
817Quantum™ Simply Cellular® anti-Rat IgGPDS 814.pdf & PDS 818SDS ST337.pdf

References

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). For additional information about working with these products, see our product data sheet on Antibody Titration or check out PDS 818, Tips and Troubleshooting.

Catalog CodeDescriptionProduct Data SheetSafety Data Sheet
810Simply Cellular® anti-Mouse IgGPDS 810.pdfSDS ST337.pdf
812Simply Cellular® anti-Human IgGPDS 812.pdfSDS ST337.pdf
813Simply Cellular® anti-Rat IgGPDS 813.pdfSDS ST337.pdf

QuickCal®

QuickCal® constructs a calibration curve associating fluorescence channel values to standardized fluorescence intensity units. The channel value corresponding to the kit's blank bead is converted to the appropriate standardized fluorescence intensity unit (MESF or ABC); this will be the instrument's detection threshold for the specific reporter and detector. For instructions regarding QuickCal, please see Product Data Sheet 819, QuickCal® v. 2.3 Data Analysis Program or watch our video.