Micromanipulation by "multiple" optical traps created by a single fast scanning trap integrated with the bilateral confocal scanning laser microscope.

We have developed a novel micromanipulator consisting of multiple optical traps created by scanning one single beam trap along a variable number of positions. Among other things, this enables the orientation of irregularly shaped and relatively large structures which could not be oriented by just one trap as is demonstrated on long Escherichia coli bacteria filaments. We expect that the multiple trap manipulator will broaden the field of applications of optical trapping as a micromanipulation technique. For example, it facilitates the study of mechanical properties of extended structures as illustrated by a "bending"-experiment using E. coli bacterium filaments. A special application of the multiple trap manipulator is the "indirect trapping" of objects which we did by keeping them held between other optically trapped particles. Indirect trapping makes it possible to trap particles which either cannot be trapped directly due to their optical properties (refractive index) or for which exposure to the laser radiation is undesirable. The multiple optical trap manipulator is controlled interactively by a UNIX workstation coupled to a VME instrumentation bus. This provides great flexibility in the control of the position and the orientation of the optical traps. Micromanipulation makes it desirable to have real time 3D microscopy for imaging and guidance of the optical traps. Therefore we integrated optical micromanipulation and a specially developed real-time confocal microscope. This so called bilateral confocal scanning laser microscope (bilateral CSLM) [Brakenhoff and Visscher, J Microsc 165:139-146, 1992] produces images at video rate.