Probing Real-Time Activation of Nf-Kappa B Signaling Circuits by Low Level Laser Therapy in Single Living Cells

The purpose of this project is to investigate the activation of NF-kappa B signaling circuits by low level laser therapy (LLLT) in single living cells using fluorescence-based technology, together with our high-resolution optical tweezers system, fluorescence detection system, microfluidic system, accurate sample temperature control system, micro-scale surface enhanced Raman spectroscopy system, laser-guided direct writing cell bioprinting system and image-based autofocusing system. This seven in one system allows us to probe the generation of reactive oxygen species (ROS) and the real-time activation of NF-kappa B signaling circuits by LLLT in single living cells. This project will primarily focus on three specific aims, and we will outline them briefly: Aim 1: Construction of precise near-infrared laser system, together with the microfluidic cell culture platform and fluorescence microscopy for LLLT; Aim 2: Probing real-time fluorescent signals of both ROS and NF-kB using two-channel, simultaneous-imaging system, and characterizing the effects of LLLT dose on ROS formation and NF-kB activation, and Aim 3: Validate whether the dysfunction of mitochondria due to the presence of either a mitochondrial uncoupler, carbonyl cyanide m-chlorophenylhydrazone (CCCP) or knockdown of cytochrome c oxidase can be naturally restored via LLLT. We anticipate the proposed LLLT in single living cells is an effective treatment strategy and LLLT can naturally restore mitochondrial function.