Figure 1.

The concept of performing single-cell multiplex RT-PCR using microcapsules. The individual cells are isolated (compartmentalized) in semi-permeable microcapsules and are lysed by dispersing microcapsules in an appropriate lysis buffer. Upon lysis, the nucleic acid molecules longer than 300 bp. are retained within the microcapsules, while low molecular weight biomolecules are removed by dialysis. Following cell lysis step, the microcapsules carrying purified nucleic acids are dispersed in RT-PCR reaction mix containing fluorescently labelled oligonucleotides. Once dispersed in a reaction mix, the reaction components quickly diffuse to the microcapsule enabling multiplex RT-PCR reaction on encapsulated nucleic acids. After RT-PCR the microcapsules carrying a particular cell type, and the transcript(s) of interest, become fluorescent thus enabling differentiation of encapsulated cells based on their digital transcriptional profile.