Typical cell images for clustered cells composed of more than three cells in cancer cell-implanted and control blood.

<p>Each data count (<i>n</i>) indicates the image number having the same cluster size and <i>N_n</i>. Bars, 20 µm.</p

Summary of total cell area, <i>S</i>, total nucleus area, <i>S_n</i>, number of nuclei, <i>N_n</i>, and perimeter ratio, <i>R</i>, for each cluster size.

<p>Summary of total cell area, <i>S</i>, total nucleus area, <i>S_n</i>, number of nuclei, <i>N_n</i>, and perimeter ratio, <i>R</i>, for each cluster size.</p

Results of quantitative gene copy number assays.

<p>(a) Results of CGH assays performed for the MAT-LyLu cell line. Liver tissue of the rat was used as a reference. Gene amplifications for <i>csrp2</i> and <i>zdhhc17</i> located on chromosome 7q13 were found. (b) Results of TaqMan copy number assays performed with clusters larger than 300 µm² collected in the collection reservoir, and cells smaller than 300 µm² collected in the discarded reservoir. Results of the assays for the MAT-LyLu cell line (positive control) and liver tissue (negative control) are also shown.</p

Summary of image processing.

<p>Firstly, photographs of both a cell and the background were taken. Next, the background image was subtracted from the cell image and holes were filled. Finally, imaging biomarkers, <i>S</i>, <i>S_n</i>, <i>N_n</i>, and <i>R</i>, were calculated. Bars, 10 µm. The hole-filling procedure is explained as indicated by an asterisk. Bar, 10 µm.</p

Typical cell images for single and double cells in cancer cell-implanted and control blood.

<p>Each data count (<i>n</i>) indicates the image number having the same cluster size and <i>N_n</i>. Bars, 20 µm.</p

Instrumental set-up.

<p>(a) Summary of the on-chip multi-imaging flow cytometry system. The system was composed of seven major modules: (i) microchip, (ii) bright-field (BF) imaging, (iii) fluorescent (FL) detection, (iv) multi-view, (v) CCD camera, (vi) sorting, and (vii) controller, as numbered in the figure. (b) Summary of the multi-view module. (c) A photograph of the system.</p

An example of cell sorting.

<p>Two photographs of the discarded reservoir (a) and the collection reservoir (b) indicated in the chip photograph are shown. Clustered cells are indicated by white arrows. Bars, 100 µm.</p

Summary of the number of nuclei, <i>N_n</i>.

<p>(a) A histogram of <i>N_n</i> obtained from cancer cell-implanted and control blood. (b) The relationship between <i>N_n</i> and cell cluster size.</p

Histograms of total cell area, S, for cancer cell-implanted (a and c) and control blood (b and d).

<p>Two threshold values (a) and (b) for cluster identifications are indicated as dotted and dashed lines.</p