Figure 2
From: High throughput single cell analysis of mitochondrial heteroplasmy in mitochondrial diseases
(A) Representative image of droplets containing a single cell. Arrowheads show nuclei of cells dyed by Hoechst 33342. (B) sc-ddPCR protocol that we used for this work. (C) Threshold line setting for the set of probes. Threshold lines were set by ddPCR using plasmids carrying a NHDF-specific (healthy) target sequence (NHDF plasmid) and mutation-specific target sequence (BK01, BK02, BK04, GM01503A, and GM03672 plasmids). (D) Results of ddPCR using plasmids for threshold line setting. (E) Threshold lines validated by the proportional relations between the cell copy number of the positive signals above the threshold and the loaded cell number. The upper figure shows the cell copy number of NHDF-specific (healthy) target sequences detected by each set of probes when encapsulating NHDFs at a concentration of 2.5 × 105/ml or 1.25 × 105/ml. The lower figure shows the cell copy number of the mutant-specific target sequence detected when encapsulating mutant cells at a concentration of 2.5 × 105/ml or 1.25 × 105/ml.
