An improved optical lock-in detection method for contrast-enhanced imaging in living cells
Synthetic and genetically encoded optical switchprobes and lock-in detection methods provide a means tomodulate and isolate fluorescence signals from the optical switchand protein complexes of interest against un-modulatedbackground sources. In this paper we propose an improvedmethod for optical lock-in detection (OLID) fluorescencemicroscopy that can significantly enhance image contrast ofoptical switch probes within living cells. In particular, weintroduce a method for automatic and optimal determination of the reference waveform, which is consequently used for thecorrelation analysis and construction of the contrast-enhancedimage. The method involves computing the average scope of eachpixel in the image over several switching cycles and then selectingthe pixel with maximum scope as a reference point. Thecorrelation between the fluorescence emission of each pixel over the switching cycles and the reference waveform is thencomputed. Finally a new contrast-enhanced image is constructedbased on the scope-weighted correlation value. Our results ofanalysis show that this method and algorithm can remarkablyimprove contrast enhancement and noise reduction compared to the original method.
Gan Du Gerard Marriott Yuling Yan
School of Engineering, Santa Clara University Santa Clara, CA, 95053, U.S.A + University of California at Berkeley, Berkeley, CA, 94720, U.S.A
国际会议
成都
英文
1-5
2010-06-18（万方平台首次上网日期，不代表论文的发表时间）