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Structured Illumination Imaging
Akin
to conventional confocal microscope imaging, this revolutionary
technology allows optical sectioning... particularly useful in
fluorescence microscopy. Fluorescence occurs when a dye absorbs a
higher-energy photon and then
emits a lower one, (e.g. blue light is absorbed and green light is
emitted). Fluorescent dyes are attached to proteins or tissue
structures
that reside along the height dimension of the sample. The problem is
that the dyes attach on various heights, therefore when fluorescent
samples are excited, emitted photons come from all height levels of the
sample. So fluorescent microscope images suffer from stray light and
hazing effect.
Various techniques exist to clean up fluorescent images, such as
laser confocal imaging and software-driven deconvolution imaging.
Structured Illumination (SI) is the latest image enhancement technique
on the scene, having been pioneered at Oxford
University and 1st patented in 1997. The technique involves imaging a
ronchi ruling onto a sample, then moving the ruling in 3 steps, each by
1/3 of the ruling’s pitch. Sample heights within the focal range of the
object show those in-focus regions with a grid image on them.
A proprietary algorithm re-combines these 3 images in a manner that
extinguishes any portion of the images that has un-focused light on
them, while eliminating the “grid-image” from the final resultant
image. This technique is substantially less costly than laser scanning
confocal or Nipkow confocal technologies.
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