Figure 2. Identification and classification of acute lymphocytic leukemia cell.

(a) Representative confocal microscopy images of RS4;11, REH and MN60 B-leukemia cells fixed and processed for immunofluorescence analysis with anti-CD38, anti-CD19, anti-CD10 and anti-CD20 monoclonal antibodies (red), to monitor their expression levels. Gray, Hoechst-33258 nucleic-acid staining. Scale bar: 10 μm. (b) Representative immunoblotting of RS4;11, REH and MN60 cells with antibodies against CD10, CD19, CD20 and CD38 (as indicated). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is shown for the internal protein levels and molecular weight standards (kDa) are indicated on the left of each panel. The blots have been run under the same experimental conditions. (c) Mean Raman spectra of normal B-lymphocytes and the three analyzed B-leukemia cell lines (as indicated). Each spectrum is an average of 300 cells. (d) Raman spectral classification and assignment²²,²³. Abbreviations: def, deformation; str, stretching; bk, vibration of backbone; sym, symmetric; A, adenine; C, cytosine; G, guanine; T, thymidine; U, uracil (ring breathing modes of DNA/RNA bases); Phe, phenylalanine; Tyr, tyrosine; Trp, tryptophan; NA, nucleic acids, P, proteins; L, lipids. (e) Difference spectra obtained by subtracting the normal B cell mean Raman spectrum from the RS4;11, REH and MN60 B-leukemia cell mean Raman spectra (as indicated). (f) PCA scatter plots comparing control B-lymphocytes and B-leukemia cells. (g) Confusion matrix for the classification of the control B-lymphocytes and the B-leukemia cells.