Optimization of optical trapping and laser interferometry in biological cells

Optical trapping and laser interferometry enable the non-invasive manipulation of colloids, which can be used to investigate the microscopic mechanics of surrounding media or bound macromolecules. For efficient trapping and precise tracking, the sample media must ideally be homogeneous and quiescent whereas such conditions are usually not satisfied in vivo in living cells. In order to investigate mechanics of the living-cell interior, we introduced (1) the in-situ calibration of optical trapping and laser interferometry, and (2) 3-D feedback control of a sample stage to stably track a colloidal particle. Investigating systematic errors that appear owing to sample heterogeneity and focal offsets of a trapping laser relative to the colloidal probe, we provide several important caveats for conducting precise optical micromanipulation in living cells. On the basis of this study, we further improved the performance of the techniques to be used in cells, by optimizing the position sensitivity of laser interferometry and the stability of the feedback simultaneously.