A combined double-tweezers and wavelength-tunable laser nanosurgery microscope

Abstract

In two previous studies we have conducted combined laser subcellular microsurgery and optical trapping on chromosomes in living cells^{1, 2}. In the latter study we used two separate microscopes, one for the trap and one for the laser scissors, thus requiring that we move the cell specimen between microscopes and relocate the irradiated cells. In the former paper we combined the 1064 nm laser trap and the 532 nm laser scissors into one microscope. However, in neither study did we have multiple traps allowing for more flexibility in application of the trapping force. In the present paper we describe a combined laser scissors and tweezers microscope that (1) has two trapping beams (both moveable via rapid scanning mirrors (FSM- 300, Newport Corp.), (2) uses a short pulsed tunable 200 fs 710-990 nm Ti:Sapphire laser for laser microsurgery, and (3) also has the option to use a 337 nm 4 ns UV laser for subcellular surgery. The two laser tweezers and either of the laser ablation beams can be used in a cell surgery experiment. The system is integrated into the robotic-controlled RoboLase system³. Experiments on mitotic chromosomes of rat kangaroo PTK2 cells are described.