Lateral modulation boosts image quality in single plane illumination fluorescence microscopy

Abstract

A new microscope combines optical sectioning by fluorophore excitation using a single light sheet with structured illumination. Several images with laterally intensity-modulated light sheets are recorded from scattering fluorescent specimens. By applying a simple data processing scheme, the nonmodulated volumes are identified. The blurred features become dark, and the resultant images are improved in terms of contrast and resolution. Hence, the instrument is capable of discriminating against contributions to the image that are induced by the optical properties of the specimen. The new microscope's capabilities are demonstrated by imaging the internals of the head of an adult Drosophila melanogaster (fruit fly) expressing green fluorescent protein-labeled polycomb proteins.

Application of structured illumination for multiple scattering suppression in planar laser imaging of dense sprays

“…By calculating the pair-wise difference between the three recorded images, most of the multiply scattered light is removed. This is described by the following equation [17]: …”

“…This technique is inspired by Structured Illumination Microscopy (SIM) [16] but is implemented here in a planar laser imaging configuration similar to the work first published by Breuninger et al known as Single Plane Illumination Microscopy -Structured Illumination (SPIM-SI) [17]. Unlike to Breuninger's application, which concerns the study of static solid objects at microscopic scale, we are imaging dynamic liquid structures from high-velocity two-phase flows at a submillimetre scale.…”

Fuel concentration in isothermal Diesel sprays through structured planar laser imaging measurements

“…Structured light has been used in fluorescence microscopy for a decade now and it demonstrated to improve image quality and back scattering detection [15]. A couple of years ago, Breuninger et al [16] applied a structured illumination together with the so called Planar Laser Imaging technique and used it to study the fluorescence spectroscopy of a biological sample.…”

“…This function of illumination would then provide a two-dimensional image of intensity I R (x, y) such as [16]:…”

Propagation stability of self-reconstructing Bessel beams enables contrast-enhanced imaging in thick media

“…Furthermore, spreading of the beam along the propagation direction leads to augmented illumination of out-of-focus objects thereby decreasing axial resolution and contrast for higher penetration depths. Approaches to remove background blur and to increase image contrast include structured illumination 8,11,12 and variations thereof 13,14 . These methods are based primarily on a removal of out-of-focus light in a post-processing step.…”