Recent developments in ﬂuorescence microscopy with speciﬁc reference to light-sheet ﬂuorescence microscopy (LSFM), multi-photon intra-vital microscopy and super-resolution microscopy are discussed in a review article by Stellenbosch University Central Analytical Facility and Department of Physiological Sciences researchers. The latest development of combining the different technologies to improve live imaging and overcoming the limitations, down to a cellular level, are also presented.
Advances in fluorescence microscopy are beneficial for research in regenerative health. Live-cell imaging is a powerful tool to understand the underlying mechanisms of tissue regeneration.
Some of the most challenging limitations in microscopy, such as slow acquisition speed, the resolution limit of light, low signal to noise ratio in thick samples, as well as photobleaching and phototoxicity have been overcome by recent advances in microscopy. In applications such as light-sheet fluorescence microscopy and intra-vital multi-photon microscopy, improved deep tissue imaging has been achieved, and super-resolution technologies have shown to improve optical resolution far beyond the diffraction limit of light for better visualisation at the cellular and molecular level. Researchers can now image live samples at a much higher resolution for a prolonged time by combining certain techniques. Advances in analytical technologies will enable researchers to gain an even better understanding of the processes involved to ultimately translate stem cell research into therapeutic interventions.