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) .



In this work we used our set up consisting of an optical tweezers plus non-linear micro-spectroscopy system to perform scanning microscopy and observe spectra using two photon excited (TPE) luminescence of captured single cells conjugated with quantum dots of US and CdTe. The US nanocrystals are obtained by our group via colloidal synthesis in aqueous medium with final pH = 7 using sodium polyphosphate as the stabilizing agent. In a second step the surface of US particles is functionalized with linking agents such as Glutaraldehyde. The CdTe quantum dots are functionalized in the its proper synthesis using mercaptoacetic acid (AMA). We used a femtosecond Ti:sapphire laser to excite the hyper Rayleigh or TPE luminescence in particles trapped with an Nd:YAG cw laser and a 30 cm monochromator equipped with a cooled back illuminated CCD to select the spectral region for imaging. With this system we obtained hyper Rayleigh and TPE luminescence images of macrophages and other samples. The results obtained show the potential presented by this system and fluorescent labels to perform spectroscopy in a living trapped microorganism in any neighbourhood. and dynamically observe the chemical reactions changes in real time.

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