Electroporation Assisted Surface-enhanced Raman Spectroscopy for Living Cells
|School||Fujian Normal University|
|Keywords||Electroporation silver nanoparticles living cells Surface Enhanced Raman Scattering (SERS)|
In this thesis, the application of surface-enhanced Raman scattering (SERS) spectroscopy technology in the detection and analysis of the living cells was presented. In current intracellular SERS measurements, gold or silver nanoparticles are delivered into living cells by "passive uptake". This procedure is time-consuming and less optimal for the real-time intracellular SERS analysis. Moreover, "passive uptake" is poor efficient for delivery of silver nanoparticles into suspension cells. To overcome the disadvantages of the existing delivery techniques, we have developed a new method based on electroporation that can rapidly deliver Au/Ag nanoparticles into living cells within one minute and successfully carried out intracellular SERS measurements. The main research as follow:1. Silver nanoparticles inside a cell were seen in the electron micrograph after electroporation and SERS spectra were successfully obtained from C666, A431 and CA46 cell lines using electroporation delivery of silver nanoparticles in less than one minute.2. We explored the effect of silver colloid concentration for fastest delivery of silver nanoparticles into living C666 cells for SERS. The viability of cells, SERS spectra, the integrated SERS intensity and the reproducibility of spectrum were compared in six electroporation experiments of the same electric pulse parameters but different concentration of silver colloid.3. We optimized the electroporation parameters, including adjustment of the pulse pattern, operation temperature, and electroporation buffer, for fastest delivery of silver nanoparticles into living C666 cells with low concentration of silver colloid.4. Since "passive uptake" is time consuming, poor efficient and non-uniform distribution for delivery of silver nanoparticles into suspension cells, our results indicated that considerable amount of silver nanoparticles were rapidly delivered into the HL60 cells (as the model system of suspension cells) by electroporation and good quality SERS spectra were obtained while the viability of the cells was maintained.As a result, the fast electroporation-SERS method can be utilized to deliver silver nanoparticles into living cells for intracellular SERS measurements. The process of sample preparation before SERS detection is accelerated significantly, demonstrating great potential for the biomedical applications of SERS, such as real time detection and analysis of the biochemical substance in living cells.