Figure 3: Expression-based CD64 cell capture in the capture chamber immobilized with anti-CD64 (clone 10.1) antibody. | Nature Communications

Figure 3: Expression-based CD64 cell capture in the capture chamber immobilized with anti-CD64 (clone 10.1) antibody.

From: A point-of-care microfluidic biochip for quantification of CD64 expression from whole blood for sepsis stratification

Figure 3

(a) Antibody adsorption characterization of the capture chamber. BSA-blocked chamber shows no fluorescence; however, PE-conjugated anti-CD64 antibody-filled chamber shows high fluorescence. In antibody-adsorbed chamber, white circular patterns at the periphery of the pillars show the adsorbed antibody after the unadsorbed antibody is flushed away. Intensity plot along A–B, with intensity peaks represent the white regions around pillars’ periphery and valleys represent pillars. (b) CD64 expression-based capture of monocytes using Sample A (mCD64=6.03), with inset CD64 histograms of monocytes before capture (Blue) and after capture (Red) shows almost complete capture. The red and blue curves show the exit and the entrance normalized percent monocyte count versus mCD64 expression,respectively, with green bars representing difference of entrance minus exit cell counts. (c) CD64 expression-based capture of neutrophils using Sample A (nCD64=2.43). (d) CD64 expression-based capture of monocytes using Sample B (mCD64=3.9). (e) CD64 expression-based capture of neutrophils using Sample B (nCD64=1.08). (f) The amplitude histogram of the cell pulses from the entrance counter (green) and exit counter (red). (g) False-coloured fluorescent image showing the captured CD64 cells around the pillars. Cells are captured specifically with antigen–antibody interaction as they lie alongside the streamlines (scale bar: 40 μm). (h) The plot shows a linear correlation between percent granulocytes + monocytes capture versus nCD64 expression ratio with (coefficient of determination: R2=0.87, P<0.0001). (i) The percent granulocytes + monocytes capture averaged from all samples in a bin versus 0.25 nCD64 bins. The plot shows a linear correlation between them with (coefficient of determination: R2=0.96, P<0.0001) with error bars representing s.d in cell capture. The green bars of secondary y axis represent the number of samples in each bin.

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