Evaluating performance in three-dimensional fluorescence microscopy

Evaluating performance in three-dimensional fluorescence microscopy

In biological fluorescence microscopy, image contrast is often degraded by a high background arising from out of focus regions of the specimen. This background can be greatly reduced or eliminated by several modes of thick specimen microscopy, including techniques such as 3-D deconvolution and confocal. There has been a great deal of interest and some confusion about which of these methods is '...

Three-dimensional holographic fluorescence microscopy.

Most commonly used methods for three-dimensional (3D) fluorescence microscopy make use of sectioning techniques that require that the object be physically scanned in a series of two-dimensional (2D) sections along the z axis. The main drawback in these approaches is the need for these sequential 2D scans. An alternative approach to fluorescence imaging in three dimensions has been developed tha...

Three dimensional image restoration in fluorescence lifetime imaging microscopy.

A microscope set-up and numerical methods are described which enable the measurement and reconstruction of three-dimensional nanosecond fluorescence lifetime images in every voxel. The frequency domain fluorescence lifetime imaging microscope (FLIM) utilizes phase detection of high-frequency modulated light by homodyne mixing on a microchannel plate image intensifier. The output signal at the i...

High-resolution fluorescence microscopy using three-dimensional structured illumination

We developed a high-resolution microscope based on three-dimensional structured illumination generated with two spatial light modulators. This setup enables both lateral resolution improvement by a factor two and axial localization of point like objects with nanometric precision.

Three - Dimensional Fluorescence Microscopy by Optical Scanning Holography