Abstract: Methods of identifying a fluorochrome panel suitable for use in a flow cytometric protocol are provided. Methods of interest include receiving an instrument identifier and a request for N fluorochrome identifiers, selecting two or more subsets of N fluorochrome identifiers from a set of fluorochrome identifiers in a spectral matrix associated with the instrument identifier, and identifying the fluorochrome panel from the two or more subsets of N fluorochrome identifiers based on a numerical stability metric calculated for each of the two or more subsets of N fluorochrome identifiers using the spectral signatures associated with the two or more subsets of N fluorochrome identifiers. Systems and non-transitory computer readable storage media for practicing the invention are also provided.

Abstract: Disclosed herein include methods, compositions, and kits suitable for spectral unmixing and compensation in multi-parameter flow cytometry. Disclosed herein include reagents which comprise two antibodies conjugated to one another to form a bispecific reagent (e.g., a bispecific probe). A first antibody can have affinity for a highly expressed antigen on the surface of a cell of interest and a second antibody can have affinity for each of the antibody-dye conjugates in a multi-parameter panel. There are provided, in some embodiments, methods of using the bispecific reagent to determine spillover, perform compensation, and generate a compensation matrix.

Abstract: Systems, methods, compositions, and kits for measuring secreted factors from cells are disclosed herein, including those capable of determining single cell secretion activity and protein expression and/or gene expression simultaneously. Disclosed herein include bispecific probes comprising an anchor probe capable of specifically binding to a surface cellular target of a cell, and a capture probe capable of specifically binding to a secreted factor secreted by a cell that is associated with the capture probe. Also disclosed herein include secreted factor-binding reagents capable of specifically binding to a secreted factor bound by a capture probe, where a secreted factor-binding reagent can comprise a secreted factor-binding reagent specific oligonucleotide comprising a unique factor identifier sequence for the secreted factor-binding reagent.

Abstract: Methods of assessing the immunomodulatory potential of a multipotent stromal cell (MSC) population are provided. Aspects of the methods include evaluating the amount of CD54/IL-6 associated with an MSC in a sample of the MSC population to obtain a CD54/IL-6 result and providing an assessment of the immunomodulatory potential of the MSC population based on the obtained CD54/IL-6 result. Also provided are systems and kits that find use in practicing the subject methods.

Abstract: Methods of assessing the immunomodulatory potential of a multipotent stromal cell (MSC) population are provided. Aspects of the methods include evaluating the amount of CD54/IL-6 associated with an MSC in a sample of the MSC population to obtain a CD54/IL-6 result and providing an assessment of the immunomodulatory potential of the MSC population based on the obtained CD54/IL-6 result. Also provided are systems and kits that find use in practicing the subject methods.

Abstract: The present invention provides a method of identifying sub-populations of cells in a cellular sample. Aspects of the method include categorizing cells of the cellular sample into at least a first and second population based on a first phenotypic property. The method may further include sub-categorizing each of the first and second population into sub-populations of cells based on a second and third phenotypic property, e.g., by using X detectable labels providing Y distinct signals, wherein X>Y, to identify sub-populations of cells in the cellular sample.