Method for analysis of cytochrome C in intact mitochondria using giant Raman spectroscopy on nanostructured coatings

摘要

1. The method of analysis of cytochrome C in intact mitochondria using giant Raman spectroscopy (GCR), including the preparation of mitochondria and their deposition on a substrate based on a dielectric chemically inert material with a nanostructured coating with a thickness of 1-10 μm in the form of ring silver nanostructures, while the rims silver rings consist of porous silver aggregates communicating with each other on the surface of which are rounded silver nanoparticles 2-90 nm in size, followed by immobilization mitochondria data on nanostructured coating, detection of SERS spectra with the subsequent decoding characteristic oscillations analysis sample SERS spectra using standard software obespecheniya.2. The method according to claim 1, characterized in that as a dielectric chemically inert material, a material selected from the series is used: aluminum oxide, magnesium oxide, silicon dioxide, zirconia, silicate glass. 3. The method according to claim 1, characterized in that the nanostructured coatings in the form of ring silver nanostructures provide effective contact with mitochondria and allow their diagnosis by the SERS method, the distance between the nanostructures and the surface of the mitochondria is not more than 20 nm. The method according to claim 1, characterized in that for the detection of cytochrome C, liquid samples of 300 microliters in volume are used by pricking them onto a nanostructured coating previously placed in a glass bottom Petri dish. A method according to claim 1, characterized in that the time of immobilization of mitochondria on nanostructured coatings is from 2 to 60 minutes. Spos