In:
Review of Scientific Instruments, AIP Publishing, Vol. 86, No. 6 ( 2015-06-01)
Abstract:
We automate the manipulation of genomic-length DNA in a nanofluidic device based on real-time analysis of fluorescence images. In our protocol, individual molecules are picked from a microchannel and stretched with pN forces using pressure driven flows. The millimeter-long DNA fragments free flowing in micro- and nanofluidics emit low fluorescence and change shape, thus challenging the image analysis for machine vision. We demonstrate a set of image processing steps that increase the intrinsically low signal-to-noise ratio associated with single-molecule fluorescence microscopy. Furthermore, we demonstrate how to estimate the length of molecules by continuous real-time image stitching and how to increase the effective resolution of a pressure controller by pulse width modulation. The sequence of image-processing steps addresses the challenges of genomic-length DNA visualization; however, they should also be general to other applications of fluorescence-based microfluidics.
Type of Medium:
Online Resource
ISSN:
0034-6748
,
1089-7623
Language:
English
Publisher:
AIP Publishing
Publication Date:
2015
detail.hit.zdb_id:
209865-9
detail.hit.zdb_id:
1472905-2
SSG:
11
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