Stimulated Raman scattering microscopy in chemistry and life science – Development, innovation, perspectives

In this review, we present a summary of the basics of the Stimulated Raman Scattering (SRS) phenomenon, methods of detecting the signal, and collection of the SRS images. We demonstrate the advantages of SRS imaging, and recent developments, but also the limitations, especially in image capture speeds and spatial resolution. We also compare the use of SRS microscopy in biological system studies with other techniques such as fluorescence microscopy, second-harmonic generation (SHG)-based microscopy, coherent anti-Stokes Raman scattering (CARS), and spontaneous Raman, and we show the compatibility of SRS-based systems with other discussed methods. The review is also focused on indicating innovations in SRS microscopy, on the background of which we present the layout and performance of our homemade setup built from commercially available elements enabling for imaging of the molecular structure of single cells over the spectral range of 800-3600 cm-1. Methods of image analysis are discussed, including machine learning methods for obtaining images of the distribution of selected molecules and for the detection of pathological lesions in tissues or malignant cells in the context of clinical diagnosis of a wide range of diseases with the use of SRS microscopy. Finally, perspectives for the development of SRS microscopy are proposed.