fig4

Figure 4. Different ways of single-cell isolation. A: Laser capture microdissection. A thermolabile polymer is placed on a tissue section on a glass slide. An infrared laser fires through the cap over the cells of interest to melt the film. The cell of interest adheres to the film, leaving the unwanted cells behind; B: fluorescence-activated cell sorting. A stream of single cells passes through an excitation laser beam and the fluorescent signal is analyzed by a multispectral detector. Single cells can be sorted into a 96 well plate; C: microfluidic-based single-cell isolation: i) An example showing a microfluidic device for single cell gene expression analysis (figure is adapted from White et al.^[94,95], 2011): (1) loading of single cells; (2) capturing single cells; (3) reverse transcription; (4) PCR; ii) Gel Bead-in-EMulsions (GEMs) formation and barcoding of 10× Genomics single-cell sequencing platform (figure is adapted from 10× Genomics Inc). Single cell GEMs are generated by passing cells with enzyme mix, partitioning oil and 10× barcoded gel beads. After GEM formation, the gel bead is dissolved and the co-partitioned cell is lysed. Reverse transcription occurs inside GEMs and barcoded full-length cDNA is generated. After RT, the GEMs are broken and the cDNA is pooled prior to library preparation for sequencing