Researchers have developed a fluorescence microscope that uses structured illumination for fast super-resolution imaging over a wide field of view. The new microscope was designed to image multiple living cells simultaneously with a very high resolution to study the effects of various drugs and mixtures of drugs on the body.
Researchers developed a fluorescence microscope that uses structured illumination for fast super-resolution imaging over a wide field of view. It can also be used for multicolor and high-speed imaging. Credit: Henning Ortkrass, Bielefeld UniversityThe new microscope is based on super-resolved structured illumination microscopy , which uses a structured pattern of light to excite fluorescence in a sample and achieve a spatial resolution beyond the diffraction limit of light.
To achieve high resolution across a wide field of view, the new microscope reconstructs super-resolved images from a set of raw images. These raw images are acquired by using a set of six optical fibers to illuminate the sample with a sinusoidal striped pattern that is shifted and rotated to gain extra information. This creates a two-fold"The fiber selection andis performed using a newly designed fiber switch based on galvanometric mirrors and MEMS-mirrors," said Ortkrass.
The researchers used their new microscopy setup to image fixed multicolor-stained liver cells. The image showed the cells’ tiny membrane structures, which are smaller than the diffraction limit of light. Credit: Henning Ortkrass, Bielefeld UniversitySince the liver is the primary organ involved in drug metabolism, the researchers tested the setup using samples of fixed multicolor-stained rat liver cells.
"This compact system uniquely combines a large field of view and fast pattern switching speed with multicolor, power-efficient excitation," said Ortkrass."In addition, the setup achieves very high image quality and can be tuned to perform either 2D-SIM or TIRF-SIM." Next, the researchers plan to apply the microscopy setup to live cell studies of liver cells to observe the dynamics of cells treated with several drugs. They also plan to improve the image reconstruction process to accomplish live reconstruction of the acquired raw data.
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