V. Richter, M. Piper, M. Wagner, H. Schneckenburger: „Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM)”, Appl. Sci. 9 (6) (2019) 1188, doi: 10.3390/app9061188.
von Herbert, Schneckenburger
In the context of various approaches for super-resolution microscopy Structured Illumination Microscopy (SIM) offers several advantages: it needs rather low light doses (with a low risk of phototoxicity or photobleaching), is comparably fast and flexible concerning the use of microscopes, objective lenses and cameras, and has a potential for 3D imaging. This paper describes an experimental setup for SIM with first diffraction orders of a spectral light modulator (SLM) creating an interference pattern in two dimensions. We kept this system rather compact with a comparably large illuminated object field, validated it with nano-beads and applied it further to living cells for imaging the cytoskeleton, mitochondria or cell nuclei with a resolution slightly above 100 nm. Its advantages, challenges and limitations – concerning cameras, acquisition time, depth of imaging, light exposure, and combination with further super-resolving methods - are discussed.