Low-NA Lens Optical Imaging for Multiplexed Tissue Characterization
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Solution Overview
Problem
Current imaging techniques for characterizing biological samples, such as cells and tissues, face challenges in maintaining spatial information, throughput, and the ability to probe multiple surface molecules simultaneously, while also preserving the integrity of the sample for downstream analyses.
Innovation Solution
An optical imaging system is developed that includes a frame for mechanical coupling between stages, a sample holding region, a low-numerical-aperture lens arrangement, and a sensor array. This system allows for the capture of fluorescent images of a sample using a substrate with microfluidic channels, enabling the sequential introduction of probe solutions and preservation of spatial information.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
If conventional immunofluorescence or immunohistochemistry is used, then spatial information is maintained, but the number of molecules that can be probed is limited to 5-7
Solution Approach 1:
The patent segments the probing process into multiple sequential rounds, where different sets of probes are applied to the same tissue section at different times. Each round targets a different subset of molecules, and the tissue is re-imaged after each probing round. This allows comprehensive molecular characterization (15-18 molecules) while preserving spatial information in the original tissue context.
Solution Approach 2:
The system employs periodic action by repeatedly applying different probe sets to the same tissue section across multiple imaging rounds. The tissue is imaged, then probed with a new set of fluorescently labeled antibodies, followed by another imaging round. This periodic probing cycle enables multiplexed molecular detection while maintaining spatial relationships.
2Adaptability or versatility
If flow cytometry or CyTOF is used, then the number of molecules probed increases to 15-18 or 100, but spatial information is lost due to single-cell analysis
Solution Approach 1:
The patent segments the analysis into sequential probing rounds applied to intact tissue sections, rather than analyzing dissociated single cells. Each round probes a specific subset of molecules while preserving tissue architecture, allowing spatial information to be maintained throughout the multiplexed molecular characterization process.
Solution Approach 2:
The system uses periodic action by cycling through multiple probing and imaging rounds on the same tissue section. Different fluorescently labeled antibody sets are applied in sequence, with imaging performed after each round, enabling detection of 15-18 molecules while maintaining spatial context that would be lost in single-cell analysis.
3Measurement precision
If conventional imaging with high numerical aperture lenses is used, then imaging resolution is improved, but the field of view is limited and cannot capture large tissue samples
Solution Approach 1:
The patent transitions from two-dimensional high-resolution imaging of small fields to three-dimensional low-resolution imaging of large fields. By accepting reduced resolution and utilizing the third dimension (multiple sequential imaging rounds), the system captures entire tissue sections at low resolution, then applies super-resolution techniques selectively to regions of interest, achieving both large field of view and high resolution where needed.
Solution Approach 2:
The imaging process is segmented into multiple rounds, where low-resolution overview images capture the entire tissue section, and high-resolution imaging is applied selectively to specific regions of interest identified in the low-resolution scans. This segmentation allows the system to manage large tissue samples efficiently while maintaining high resolution where necessary.
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
The system achieves high-resolution imaging of large tissue samples with maintained spatial information, allows for the characterization of an unlimited number of surface molecules, and preserves the sample integrity for further analyses, thereby overcoming the limitations of existing techniques.
Implementation Method 1
receiving light fluorescing from the sample at a lens arrangement disposed beneath the stage
Data Source
AI summary
An optical imaging system includes a frame designed to provide mechanical coupling between a first stage and a second stage, a sample holding region located on the first stage, a lens arrangement, and a sensor array. The lens arrangement is disposed between the first stage and the second stage and is designed to receive light from a sample at the sample holding region on the first stage. The lens arrangement has a numerical aperture less than 0.1. The sensor array is coupled to the second stage and is designed to receive light passing through the lens arrangement.