Cell-by-Cell Analysis

Fluorescent plate readers provide aggregate assay results for each well, which reflect the average characteristics of the population. Relevant subpopulations, which may be of particular importance to the subject under investigation, could be masked. In contrast, the Accuri C6 provides cell-by-cell statistics that allow differentiation between cell subpopulations. This makes it possible to derive information about the physical and chemical nature of each individual cell or particle.

Add our powerful, cost-effective cell-analysis tool to your list of essential laboratory instruments. Designed specifically with the life scientist in mind and priced comparably to a fluorescent plate reader, the Accuri C6 has a small footprint and no special set-up requirements.

Western Blots

Western blots are the gold standard for protein biomarker determination. However in many cases, flow cytometry offers specific advantages in characterizing protein expression. While Western blots represent expression of a cell population, flow cytometry characterizes individual cells, allowing further characterization of a specific cell population. This is particularly beneficial when attempting to characterize small subset cell populations.

(A) Density plot of Jurkat T cells 48 h post-transfection with either a non-specific control or Lck-specific pool of siRNA. Forward scatter (FSC-H) is on the X-axis and intracellular Lck staining on the Y-axis. A representative gate circumscribing the Lcklo subpopulation is shown. Histograms of the corresponding cell populations for Lck are shown in the insets (green histograms). Red histogram plots represent background staining with purified rabbit gamma globulin. These plots are representative of ten independent experiments.¹

RNAi Knockdown

RNAi is a powerful, genome-wide technique for studying protein loss-of-function via gene knockdown. In practice, siRNA transfection efficiencies can be low and variable, so a better approach is flow cytometric measurement of intracellular protein knockdown in situ with viable intact cells from all subpopulations. Since flow cytometry is multi-parametric, fluorescent antibodies can be used to not only assess the siRNA-targeted protein levels, but also to detect other functional cell surface marker signals in the same cell.

¹ Chan SM, Olson JA, and Utz PJ (2005) Single-Cell Analysis of siRNA-Mediated Gene Silencing Using Multiparameter Flow Cytometry. Cytometry 69A:59–65.

Download our White Paper, The Essential Cell Analysis Tool, and read more about molecular and cellular applications that take advantage of the Accuri C6’s cell analysis capabilities.

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