On-chip single cell whole genome amplification.

<p>(A) bright-field microscope image of a single SKBR-3 cell isolated in a microfluidic chamber, merged with a fluorescence picture. (B) fluorescence image of a single SKBR-3 cell stained with CellTracker Orange in the microfluidic chamber. (C and F) qPCR targeting GAPDH and ERBB2 genes of on-chip amplified DNA for quantification and characterization using 1 µl of on-chip amplified samples (5 µl) from individual SKBR-3 (n = 6) and MCF-7 (n = 6) as templates. (C) Real-time qPCR curves of GAPDH gene amplification. Amplified DNA of single SKBR-3 is represented using black curves while that of single MCF-7 cells is represented as green curves. Threshold line is in grey. (D and E) Images of fluorescence in situ hybridization (FISH) with ERBB2 probes (red) and centromere of chromosome 17 probes (green) for individual SKBR-3 (D) and MCF-7 (E). Nuclei were stained with Hoechst (blue) (F) Cq values of ERBB2 gene in a box plot. Cq values of ERBB2 were represented as black box in SKBR-3 (n = 6) and red box in MCF-7 (n = 5, * Cq of 1 sample is missing because of PCR error). (25%–75%: □, Median value:–, Mean value:□, Min/Max:°, Range of Min/Max:I).