ABSTRACT
Emerging advances in live-cell imaging have bridged gaps between biochemical, genomic, and developmental biology approaches, providing critical insights into the molecular dynamics and structure inside living cells. When choosing a microscopy system for imaging live cells four things should be considered: sensitivity of detection, speed of acquisition, resolution, and specimen viability. The main technique used in light microscopy for the past three centuries, light transmission brightfield microscopy, relies on the changes in light absorption. Widefield fluorescence microscopy (WFM) systems can be faster and more sensitive than laser scanning confocal microscopy (LSCM) systems, as light that is rejected by the pinhole on a confocal microscope is collected on a widefield microscope. The spinning disk confocal microscope is a multipoint LSCM, which captures dynamic events in a wide spectrum of timescales. Multiphoton confocal laser-scanning microscopes (MP-CLSM) present unique advantages for intravital imaging in thick tissues, in live animals or in tissue-slice culture.
