The 2014 Nobel Prize in Chemistry has been awarded jointly to William E. Moerner, Stefan W. Hell, and Eric Betzig "for the development of super-resolved fluorescence microscopy." I discuss the contributions made by this year's awardees and how advances in understanding the behavior of fluorophores and research in light microscopy converged to allow the improved visualization of biological struc...

New light microscopy techniques are pushing the limits of resolution to 50 nm and below. Fluorescence microscopy that rivals electron microscopy in resolution but operates on intact cells may be within reach.

We present Instantaneous Spatial Light Interference Microscopy (iSLIM) as a new quantitative phase method that combines the benefits of white light illumination in Zernike's phase contrast microscopy and phase stability associated diffraction phase microscopy. iSLIM is implemented as an add-on module to a commercial phase contrast microscope, and enables new features to quantitative phase imagi...

Cryogenic fluorescent light microscopy of flash-frozen cells stands out by artifact-free fixation and very little photobleaching of the fluorophores used. To attain the highest level of resolution, aberration-free immersion objectives with accurately matched immersion media are required, but both do not exist for imaging below the glass-transition temperature of water. Here, we resolve this cha...

Introduction • Methods

By combining a high-resolution image from a standard camera with a low-resolution wavefront measurement from a Shack-Hartmann sensor, we numerically reconstruct a highresolution light field. We experimentally demonstrate the method with a commercially available microscope. ©2013 Optical Society of America OCIS codes: (110.0180) Microscopy; (110.1758) Computational imaging; (100.6640) Superresol...

The invention of two-photon fluorescence light microscopy by Denk, Webb and co-workers (Denk et al., 1990) revolutionized three-dimensional (3D) in vivo imaging of cells and tissues. In 1931, the theoretical basis of twophoton excitation was established by Maria GöppertMayer, and this photophysical effect was verified experimentally by Kaiser and Garret in 1963. Two-photon excitation is a fluor...

Selective-plane illumination microscopy has proven to be a powerful imaging technique due to its unsurpassed acquisition speed and gentle optical sectioning. However, even in the case of multiview imaging techniques that illuminate and image the sample from multiple directions, light scattering inside tissues often severely impairs image contrast. Here we combine multiview light-sheet imaging w...

Optical microscopes, which are among the oldest instruments of scientific discovery, continue to be key tools in both biomedical research and routine diagnosis. This remains true despite the development of a wide range of new imaging technologies, many with far greater resolution—ranging from electron microscopes to the multitude of scanning probe systems available today. The overall simplicity...

Quantitative phase imaging (QPI) techniques are very advantageous compared to the qualitative phase imaging techniques like bright field microscopy, phase contrast microscopy, differential interference contrast microscopy due to measurement of phase information quantitatively. In QPI techniques, there is no need of exogenous contrast agents to stain or tag the specimen. White light diffraction ...