Confocal fluorescence microscopy and atomic force microscopy (AFM) are two established techniques that can be used to study single molecules. Both techniques have their individual capabilities that permit to study different aspects such as topography using AFMs, or dynamics using fluorescence spectroscopy. Thus, a combination of both techniques into a single instrument enhances the methodology ...

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...

Two-photon fluorescence microscopy is one of the most important recent inventions in biological imaging. This technology enables noninvasive study of biological specimens in three dimensions with submicrometer resolution. Two-photon excitation of fluorophores results from the simultaneous absorption of two photons. This excitation process has a number of unique advantages, such as reduced speci...

We present an imaging system that collects hyperspectral images of cells travelling through a microfluidic channel. Using a single monochrome camera and a linear variable bandpass filter (LVF), the system captures a bright field image and a set of hyperspectral fluorescence images for each cell. While the bandwidth of the LVF is 20 nm, we have demonstrated that we can determine the peak wavelen...

Spontaneous emission of fluorophores located close to a reflecting surface is modified by the interference between direct and reflected waves. The spectral patterns of fluorescent emission near reflecting surfaces can be precisely described with a classical model that considers the relative intensity and polarization state of direct and reflected waves depending on dipole orientation. An algori...