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. 2016 Mar 11;6:22975. doi: 10.1038/srep22975

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(a) Reverse transcription of miRNA based on polyA tailing method. (b) Detailed scheme for accumulation of amplicons in the LEM-PIN during PCR. (c) Comparison between the conventional and LEM-PIN qPCRs. The Ct value of the particle qPCR for PTC was increased by 3.97 with standard deviation of 0.28 compared to that of the conventional qPCR. Interestingly, particle qPCR showed no signals for NTC even up to 50^th cycle while the conventional qPCR recorded a Ct value for NTC around the 30^th cycle. (d) qPCR results for a 10-fold serial dilution of miR-9-3p. The graphs kept the uniform distances among them which are about 3.3 corresponding to the 10-fold difference in template concentration. Inset shows the standard curve drawn based on serial dilution results, which indicate the reliability of the particle qPCR recorded a PCR efficiency of more than 95%. (e) Snapshots of particle qPCR with the concentration variation of templates. The particles became fluorescent in sequence according to the concentration of template. scalebar is 1 mm. (f) Rinsing effect of the particle. Insufficiently rinsed particle (upper) showed dim fluorescence around it while perfectly rinsed one has clear contrast from the background. This could lead to the influence to the other surrounding particles when multiplexing. Scalebar is 500 μm.