Selective study of the electron transport chain components in living mitochondria is essential for fundamental biophysical research and for the development of new medical diagnostic methods. However, many important details of inter- and intramembrane mitochondrial processes have remained in shadow due to the lack of non-invasive techniques. Here we suggest a novel label-free approach based on the surface-enhanced Raman spectroscopy (SERS) to monitor the redox state and conformation of cytochrome c in the electron transport chain in living mitochondria. We demonstrate that SERS spectra of living mitochondria placed on hierarchically structured silver-ring substrates provide exclusive information about cytochrome c behavior under modulation of inner mitochondrial membrane potential, proton gradient and the activity of ATP-synthetase. Mathematical simulation explains the observed enhancement of Raman scattering due to high concentration of electric near-field and large contact area between mitochondria and nanostructured surfaces.

This paper presents a nonivasive approach to study redox state of reduced cytochromes , and of complexes II and III in mitochondria of live cardiomyocytes by means of Raman microspectroscopy. For the first time with the proposed approach we perform studies of rod- and round-shaped cardiomyocytes, representing different morphological and functional states. Raman mapping and cluster analysis reveal that these cardiomyocytes differ in the amounts of reduced cytochromes , and . The rod-shaped cardiomyocytes possess uneven distribution of reduced cytochromes , and in cell center and periphery. Moreover, by means of Raman spectroscopy we demonstrated the decrease in the relative amounts of reduced cytochromes , and in the rod-shaped cardiomyocytes caused by H₂O₂-induced oxidative stress before any visible changes. Results of Raman mapping and time-dependent study of reduced cytochromes of complexes II and III and cytochrome in cardiomyocytes are in a good agreement with our fluorescence indicator studies and other published data.

Nowadays, there is an interest in biomedical and nanobiotechnological studies, such as studies on carotenoids as antioxidants and studies on molecular markers for cardiovascular, endocrine, and oncological diseases. Moreover, interest in industrial production of microalgal biomass for biofuels and bioproducts has stimulated studies on microalgal physiology and mechanisms of synthesis and accumulation of valuable biomolecules in algal cells. Biomolecules such as neutral lipids and carotenoids are being actively explored by the biotechnology community. Raman spectroscopy (RS) has become an important tool for researchers to understand biological processes at the cellular level in medicine and biotechnology. This review provides a brief analysis of existing studies on the application of RS for investigation of biological, medical, analytical, photosynthetic, and algal research, particularly to understand how the technique can be used for lipids, carotenoids, and cellular research. First, the review article shows the main applications of the modified Raman spectroscopy in medicine and biotechnology. Research works in the field of medicine and biotechnology are analysed in terms of showing the common connections of some studies as caretenoids and lipids. Second, this article summarises some of the recent advances in Raman microspectroscopy applications in areas related to microalgal detection. Strategies based on Raman spectroscopy provide potential for biochemical-composition analysis and imaging of living microalgal cells, in situ and in vivo. Finally, current approaches used in the papers presented show the advantages, perspectives, and other essential specifics of the method applied to plants and other species/objects.

Surface-enhanced Raman spectroscopy (SERS) of living cells has rapidly become a powerful trend in biomedical diagnostics. It is a common belief that highly ordered, artificially engineered substrates are the best future decision in this field. This paper, however, describes an alternative successful solution, a new effortless chemical approach to the design of nanostructured silver and heterometallic continuous coatings with a stochastic “coffee ring” morphology. The coatings are formed from an ultrasonic mist of aqueous diamminesilver hydroxide, free of reducing agents and nonvolatile pollutants, under mild conditions, at about 200–270 °C in air. They consist of 30–100 micrometer wide and 100–400 nm high silver rings composed, in turn, of a porous silver matrix with 10–50 nm silver grains decorating the sponge. This hierarchic structure originates from ultrasonic droplet evaporation, contact-line motion, silver(I) oxide decomposition and evolution of a growing ensemble of silver rings. The fabricated substrates are a remarkable example of a new scalable and low cost material suitable for SERS analyses of living cells. They evoke no hemolysis and reduce erythrocyte lateral mobility due to suitable “coffee ring” sizes and a tight contact with the silver nanostructure. A high SERS enhancement, characteristic of pure silver rings, made it possible to record Raman scattering spectra from submembrane hemoglobin in its natural cellular environment inside single living erythrocytes, thus making the substrates promising for various biosensor chips.