Linear and non-linear micro-spectroscopy in an optical tweezers system

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(A. Periasamy & P. T. C. So, Eds.) Multiphoton Microscopy in the Biomedical Sciences V. 1000 20TH ST, PO BOX 10, BELLINGHAM, WA 98227-0010 USA: SPIE-INT SOC OPTICAL ENGINEERING (2005) .



We developed a set up consisting of an Optical Tweezers plus linear and non-linear micro-spectroscopy system to add the capabilities of manipulation and analysing the captured object. For the confocal micro-spectrometer we used a 30 cm monochromator equipped with a cooled back illuminated CCD. The spectroscopic laser system included a cw and a femtosecond Ti:sapphire lasers that allowed us to perfom raman, hyper-raman, hyper-rayleigh and two-photon excited (TPE) luminescence in trapped particles with an Nd:YAG cw laser. With the cw Ti:sapphire laser we obtained raman spectra of a single trapped polystyrene microsphere and red blood cells and silicon to evaluate the performance of our system. The femtosecond Ti:sapphire laser was used to observed hyper-rayleigh and hyper-raman peaks of SrTiO3 with 60s integration time only. In the past, hyper-raman measurements required integration times of few hours, but the huge intensity together with the 80 NMz repetition rate of the femtosecond laser decreased this time for the seconds range. The sensitiveness of our system also permitted to observe more than 14 Mie resonance peaks in the TPE luminescence of a single stained trapped microsphere, which agrees well with the calculations. This system opens up the possibility to perform spectroscopy in a living trapped micro-organism in any desired neighbourhood and dynamically observe the chemical reactions and/or mechanical properties change in real time.

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