Researchers Develop High-Speed Super-Resolution Microscope With a Large Field of View

Fluorescence Microscope

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Researchers established a fluorescence microscopic lense that utilizes structured lighting for quick super-resolution imaging over a broad field of vision. It can likewise be utilized for multicolor and high-speed imaging. Credit: Henning Ortkrass, Bielefeld University

Microscope based upon fiberoptic parts developed for studying cellular impacts of drug mixes.

Researchers have actually established a fluorescence microscopic lense that utilizes structured lighting for quick super-resolution imaging over a broad field of vision. This advanced microscopic lense is customized to catch high-resolution pictures of numerous living cells at the same time, helping with the analysis of how various drugs and their mixes affect the body.

“Polypharmacy — the effect of the many combinations of drugs typically prescribed to the chronically sick or elderly — can lead to dangerous interactions and is becoming a major issue,” stated Henning Ortkrass from Bielefeld University inGermany “We developed this microscope as part of the EIC Pathfinder OpenProject DeLIVERy, which aims to develop a platform that can investigate polypharmacy in individual patients.”

New Microscopy Liver Cell

The scientists utilized their brand-new microscopy setup to image repaired multicolor-stained liver cells. The image revealed the cells’ small membrane structures, which are smaller sized than the diffraction limitation of light. Credit: Henning Ortkrass, Bielefeld University

In the Optica Publishing Group journal Optics Express, the scientists explain their brand-new microscopic lense which utilizes fiber optics shipment of excitation light to make it possible for really high image quality over a large field of vision with multicolor and high-speed ability. They reveal that the instrument can be utilized to image liver cells, accomplishing a field of vision as much as 150 x 150 μm ² and imaging rates as much as 44 Hz while preserving a spatiotemporal resolution of less than 100 nm.

“With this new microscope, individual drug combinations can be tested on isolated cells and then imaged with super-resolution to observe dynamics of cell membrane features or organelles,” statedOrtkrass “The large field of view can provide statistical information about the cell response, which could be used to improve personalized healthcare. Thanks to the system’s potentially small size, it might also be useful for clinical applications where high resolution is important.”

New Fluorescence Microscope Setup

The brand-new fluorescence microscopic lense utilizes structured lighting for quick super-resolution imaging over a broad field of vision. Multicolor imaging can likewise be carried out, as displayed in the video. Credit: Henning Ortkrass, Bielefeld University

High resolution throughout a big field of vision

The brand-new microscopic lense is based upon super-resolved structured lighting microscopy (SR-SIM), which utilizes a structured pattern of light to thrill fluorescence in a sample and attain a spatial resolution beyond the diffraction limitation of light. SR-SIM is especially well matched for live cell imaging due to the fact that it utilizes low-power excitation that does not hurt the sample while producing extremely in-depth images.

To attain high resolution throughout a broad field of vision, the brand-new microscopic lense rebuilds super-resolved images from a set of raw images. These raw images are obtained by utilizing a set of 6 fiber optics to light up the sample with a sinusoidal striped pattern that is moved and turned to acquire additional info. This develops a two-fold resolution enhancement while still accomplishing quick imaging and working with live-cell imaging.

Super resolution Image of Multiple Cells at Once

Thanks to the microscopic lense’s big field of vision, it is possible to obtain super-resolution pictures of several cells at the same time. Credit: Henning Ortkrass, Bielefeld University

“The fiber selection and phase shift is performed using a newly designed fiber switch based on galvanometric mirrors and MEMS-mirrors,” statedOrtkrass “We also custom-designed a hexagonal holder that collimates and refocuses the beams of the six fibers into the microscope to illuminate a large FOV and allow precise adjustment of all beams. This allows the setup to be used for total internal reflection fluorescence excitation (TIRF)-SIM, which is used to restrict fluorescence excitation and detection to a thin region of the sample.”

Imaging liver cells

Since the liver is the main organ associated with drug metabolic process, the scientists checked the setup utilizing samples of repaired multicolor-stained rat liver cells. The rebuilded images produced with the brand-new microscopic lense enabled visualization of the small membrane structures that are smaller sized than the diffraction limitation of light.

“This compact system uniquely combines a large field of view and fast pattern switching speed with multicolor, power-efficient excitation,” statedOrtkrass “In addition, the setup achieves very high image quality and can be tuned to perform either 2D-SIM or TIRF-SIM.”

Next, the scientists prepare to use the microscopy setup to live cell research studies of liver cells to observe the characteristics of cells treated with numerous drugs. They likewise prepare to enhance the image restoration procedure to achieve live restoration of the obtained raw information.

Reference: “High-speed TIRF and 2D super-resolution structured illumination microscopy with a large field of view based on fiber optic components” by Henning Ortkrass, Jasmin Sch ürstedt, Gerd Wiebusch, Karolina Szafranska, Peter McCourt and Thomas Huser, 16 August 2023, Optics Express
DOI: 10.1364/ OE.495353