TY - GEN
T1 - Single molecule diffusion coefficient estimation by image analysis of simulated CCD images to aid high-throughput screening
AU - Song, Pengfei
AU - Davis, Lloyd M.
AU - Bashford, Gregory R.
PY - 2009
Y1 - 2009
N2 - Extension of one-dimensional signal analysis to two-dimensional image analysis could accelerate conventional methods of high-throughput screening in the discovery of new pharmaceutical agents. This work describes a first step taken towards this goal - the evaluation of image-analysis based estimation strategies of the diffusion coefficient of a single molecule transported within a microfabricated flowcell. A computer simulation of single-molecule imaging by a charge-coupled device (CCD) camera is used to determine if it is possible to distinguish three different types of molecules with different diffusion coefficients. The Gaussian fitting algorithm finds the variance of the transverse trajectory, which increases linearly with the diffusion coefficient; the path analysis algorithm determines the diffusion coefficient from cumulative summation of the squared displacement along the imaged path; the detector area analysis algorithm determines the number of resolvable positions or pixels in the imaged trajectory. Of the three methods, the path analysis strategy appears to provide the most reliable measure of diffusion coefficient with relative error of 13.6% and 6.4% between single molecules with diffusion coefficients of 2.85e-7 and 1.425e-7 cm2/s. The detector area analysis method can statistically distinguish between single molecules with diffusion coefficients of 5.7e-7 and 1.425e-7 cm2/s at the p0.05 level.
AB - Extension of one-dimensional signal analysis to two-dimensional image analysis could accelerate conventional methods of high-throughput screening in the discovery of new pharmaceutical agents. This work describes a first step taken towards this goal - the evaluation of image-analysis based estimation strategies of the diffusion coefficient of a single molecule transported within a microfabricated flowcell. A computer simulation of single-molecule imaging by a charge-coupled device (CCD) camera is used to determine if it is possible to distinguish three different types of molecules with different diffusion coefficients. The Gaussian fitting algorithm finds the variance of the transverse trajectory, which increases linearly with the diffusion coefficient; the path analysis algorithm determines the diffusion coefficient from cumulative summation of the squared displacement along the imaged path; the detector area analysis algorithm determines the number of resolvable positions or pixels in the imaged trajectory. Of the three methods, the path analysis strategy appears to provide the most reliable measure of diffusion coefficient with relative error of 13.6% and 6.4% between single molecules with diffusion coefficients of 2.85e-7 and 1.425e-7 cm2/s. The detector area analysis method can statistically distinguish between single molecules with diffusion coefficients of 5.7e-7 and 1.425e-7 cm2/s at the p0.05 level.
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U2 - 10.1109/IEMBS.2009.5334157
DO - 10.1109/IEMBS.2009.5334157
M3 - Conference contribution
C2 - 19964522
AN - SCOPUS:77950993151
SN - 9781424432967
T3 - Proceedings of the 31st Annual International Conference of the IEEE Engineering in Medicine and Biology Society: Engineering the Future of Biomedicine, EMBC 2009
SP - 1396
EP - 1399
BT - Proceedings of the 31st Annual International Conference of the IEEE Engineering in Medicine and Biology Society
PB - IEEE Computer Society
T2 - 31st Annual International Conference of the IEEE Engineering in Medicine and Biology Society: Engineering the Future of Biomedicine, EMBC 2009
Y2 - 2 September 2009 through 6 September 2009
ER -