Abstract
Quantitative research of cellular dynamics has been attracting more and more interest for decades. This quantitative approach promotes the research on activities at cell level, shedding light on disease therapy and intrinsic cell mechanism. In order to achieve the quantitative analysis of cell dynamics, live cell techniques, multi-dimensional measurements and efficient image analyses are the main premises. Focus drift and automatic control are the two critical aspects affecting the performance of multi-dimensional observation. Here a novel RII autofocus system is presented to solve the focus drift problem, which improves the focus resolution to 1.3-5.3% of the objective DOF. An automatic controlled spatial temporal continuous microscopy is successfully established in this study to achieve quantitative and multi-dimensional observation of live cells. Image sequences are obtained from the time lapse observation. Improved algorithms concerning background correction, segmentation and tracking were implemented efficiently to extract valuable biomedical information. The average detection rate of segmentation under different cell densities is as high as 98%. In addition, optical modulation methods are implemented as add-on components to realize specified biomedical research.