Figure 1.
Analysis of mRNA expression of thirteen B cell specific antigens in seven different cell populations isolated from peripheral blood.
For each antigen the columns from left to right correspond to CD4+ T-cells (dark gray), CD8+ T-cells (light gray), NK cells (white), CD19+ B-cells (black), CD14+ monocytes (light gray), CD33+ neutrophils (gray), and eosinophils (dark grey).
Figure 2.
Correlation of flow cytometric and nCounter results.
A: Correlation between results from the nCounter technology and flow cytometry for four typical antigens. Flow cytometric results are given as percentage of cells positive for a given antigen, and nCounter results as digital counts (after normalization and background subtraction). The number of depicted data points varies depending on the antigen tested and on the availability of both cytometric and nCounter values for a given sample. Only samples for which both values were measured are shown. Some samples, particularly informative for a particular antigen, were highlighted with dots in different shades and patterns. nBM, normal bone marrow; corr coeff, Pearson’s correlation coefficient. corr coeff >0.393 are statistically significant with p<0.01. B: Correlation between % of leukemic blasts as determined by flow cytometry, and nCounter values for two typical antigens (CD34, CD19).
Table 1.
Correlation coefficient for flow cytometry (protein) and nCounter (mRNA) values from 22 antigens.
Figure 3.
Comparative analysis of leukemia patients.
A: Comparison of nCounter values from normal BM ( = the mean of the values from eleven normal BM samples) with a sample from a patient with acute myeloid leukemia. Values in the acute leukemia sample, which were above the normal BM mean +2SD, were considered positive (light rectangles) and the corresponding mRNA expressed by the leukemic blasts. B: Correlation between % of blasts present in a leukemic sample and concordance between flow cytometry and nCounter results in forty-six patients.
Table 2.
Analysis of leukemic blasts: correlation between results from flow cytometry and nCounter analysis for twenty-two different antigens, each studied in forty-five leukemia patients.
Figure 4.
A typical case of B-ALL analyzed in parallel by flow cytometry and by the nCounter technique.
Flow cytometry shows the blast population being CD34pos, CD19pos, cy CD79a pos, cy IgM pos, TDT pos, CD10 pos and CD45 pos. In red: antigens found positive with both methods.