Single-molecule fluorescence detection using one-dimension interference pattern illumination

Single-molecule fluorescence detection using one-dimension interference pattern illumination
Interference; Illumination; Fluorescence; Nanochannel; 간섭; 조명; 형광; 나노채널
Issue Date
대한기계학회 마이크로/나노 분과
Genome wide association study requires dense maps of single nucleotide polymorphism(SNP) that cover the human genome to look for allele-frequency differences between cases and controls. Haplotype information is essential for complete description and interpretation of genomes. Despite of technical advances both in sequencing and in genotyping, haplotype information, sets of SNPs at multiple loci in chromosomes, is still missing. Because one can exactly find hidden mutants with haplotype information, cost-effective haplotyping of individuals should be available in order to advance to personalized medicine. High throughput and ultra-fast haplotyping requires accurate phasing capability, high resolving power, and highly parallelized detection schemes. In order to achieve the spatial resolution of a few hundreds nanometers, we create interference pattern illumination volumes, which is perpendicular to flow direction within nanochannel array. Currently, our system can create interferometric illumination volume with the line width of ~500 ㎚. An interference pattern illumination, with the line width of Lw = 10 ㎛, and the period of Lp = 200 ㎛, was produced as microfilm mask. DNA samples were prepared as λ-DNA stained with YOYO-1 in 1×TE buffer. Qdot605 was labeled on the λ-DNA from PNA labeling method. Single-molecule fluorescence profile of Qdot605 was measured using interference pattern illumination.
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