Multi-Spectral Structured Illumination Microscopy Parallel Acquisition
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Solution Overview
Problem
Current microscopy technologies, such as widefield and confocal microscopy, face challenges in achieving high plex and short turnaround times, while structured illumination microscopy (SIM) suffers from low throughput due to the need for multiple exposures for each frame and color channel, limiting its ability to produce high-resolution images at high imaging speeds.
Innovation Solution
The mosaic structured illumination microscope system employs a spectrally multiplexed excitation grid, using multiple colors or light sources to parallelize the imaging process, incorporating a light source, array mask, dispersion element, and sensors to achieve simultaneous imaging and demultiplexed readout, enabling faster imaging rates and higher resolution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If structured illumination microscopy is used to increase 3-dimensional resolution, then manufacturing precision is improved, but productivity deteriorates due to multiple exposures required for every frame and color channel
Solution Approach 1:
The system segments the illumination spectrum into multiple wavelength bands, with each band projected through a separate aperture of the array mask. This allows parallel acquisition of multiple wavelength channels simultaneously, eliminating the need for sequential exposures and thereby improving productivity while maintaining super-resolution capability
Solution Approach 2:
The invention adds a spectral dimension to the spatial illumination pattern by using multi-wavelength light sources. Different wavelengths are assigned to different apertures of the array mask, creating a multi-dimensional patterned illumination that enables simultaneous capture of multiple color channels, thus resolving the contradiction between resolution and acquisition speed
2Measurement precision
If multiple exposures are acquired for every frame and color channel in SIM, then measurement precision is improved, but loss of time increases
Solution Approach 1:
The system maintains continuous useful action by simultaneously acquiring multiple wavelength channels in parallel through the array mask's multiple apertures. This eliminates the sequential time loss inherent in traditional SIM, allowing continuous imaging without the time penalty of multiple sequential exposures while preserving measurement precision
3Productivity
If standard widefield microscopy is used, then productivity is maintained, but manufacturing precision deteriorates in terms of resolution
Solution Approach 1:
The system employs periodic modulation of the illumination light through the array mask apertures, creating time-varying patterned illumination at different wavelengths. This periodic action enables super-resolution imaging by encoding spatial frequency information, achieving enhanced manufacturing precision while maintaining high productivity through parallel acquisition
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach results in a 20% faster imaging rate compared to standard widefield microscopy and up to three times faster than traditional multifocal SIM, with improved 3D resolving capability and large field-of-view imaging, achieving data capture rates of 4.5 E+06 voxels per second and 3.5 emitters per cubic micron.
Implementation Method 1
a dispersion element configured to shift the first patterned light beam laterally based on the first wavelength and the second patterned light beam laterally based on the second wavelength
Implementation Method 2
an array mask having a set of apertures configured to change the first light beam to a first patterned light beam and change the second light beam to a second patterned light beam
Implementation Method 3
at least one sensor configured to detect first fluorescent radiation emitted from a sample excited by the first patterned light beam and second fluorescent radiation emitted from the sample excited by the second patterned light beam
Data Source
AI summary
In some embodiments, a system includes a light source configured to emit a first light beam having a first wavelength and a second light beam having a second wavelength. The system further includes an array mask having a set of apertures configured to change the first light beam to a first patterned light beam and change the second light beam to a second patterned light beam. The system includes a dispersion element configured to shift the first patterned light beam laterally based on the first wavelength and the second patterned light beam laterally based on the second wavelength. The system includes at least one sensor configured to detect first fluorescent radiation emitted from a sample excited by the first patterned light beam and second fluorescent radiation emitted from the sample excited by the second patterned light beam.