Fluorescence Microscopy ROI Prescan to Limit Sample Light Exposure
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Solution Overview
Problem
Conventional fluorescence microscopy methods require manual search and evaluation of samples, leading to wasted time and potential specimen damage due to excessive light exposure, which can cause photobleaching and phototoxic effects.
Innovation Solution
A method and arrangement that uses transmitted light imaging for a pre-scan to generate a stack of widefield images, allowing automated selection of regions of interest, followed by fluorescence imaging with reduced light exposure using lightsheet microscopy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional fluorescence microscopy is used to image samples, then fluorescence images can be obtained, but excessive light exposure causes photobleaching and phototoxic effects
Solution Approach 1:
The system performs transmitted light imaging as a preliminary step before fluorescence imaging to identify regions of interest. This preliminary action allows the system to pre-select which samples warrant fluorescence imaging, thereby reducing overall light exposure and preventing photobleaching and phototoxic effects while maintaining measurement precision for selected samples.
Solution Approach 2:
The system applies different imaging modalities to different regions: transmitted light imaging for initial screening of all samples, and fluorescence imaging only for selected regions of interest. This local differentiation optimizes resource allocation and minimizes harmful light exposure to samples that do not require intensive fluorescence imaging.
2Ease of operation
If manual search and evaluation of samples is performed, then sample selection can be made, but valuable time is wasted
Solution Approach 1:
The system replaces manual mechanical search and evaluation with an automated optical system that uses transmitted light imaging to automatically identify and select regions of interest. This substitution eliminates time-consuming manual operations while maintaining effective sample selection capability through automated image processing and analysis.
Solution Approach 2:
The system performs self-service by automatically analyzing transmitted light images to identify regions of interest without requiring manual intervention. The automated selection process enables the system to independently determine which samples require further fluorescence imaging, significantly reducing time loss while preserving operational effectiveness.
3Loss of information
If fluorescence imaging is performed on all samples, then comprehensive data can be obtained, but excessive light exposure damages specimens
Solution Approach 1:
The system performs transmitted light imaging as a preliminary screening step to identify which samples contain regions of interest worthy of fluorescence imaging. This preliminary action enables selective fluorescence imaging only of relevant samples, preserving comprehensive data for selected samples while avoiding unnecessary light exposure to other samples and preventing specimen damage.
Solution Approach 2:
The system applies partial action by performing fluorescence imaging only on selected regions of interest rather than on all samples. This selective approach obtains sufficient information for meaningful analysis while minimizing overall light exposure and preventing specimen damage that would result from exhaustive imaging of all samples.
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
Reduces light exposure and specimen damage by using a sample-friendly pre-scan, enabling efficient and non-destructive examination of multiple samples with improved time efficiency and accuracy.
Implementation Method 1
optics configured to selectively image the sample region in a transmitted light imaging mode or a fluorescence imaging mode. The transmitted light imaging mode is adapted to generate and process a stack of widefield images of the sample region
Implementation Method 2
The fluorescence imaging mode is adapted to obtain fluorescence images of at least one of the plurality of samples at one or more fluorescence imaging positions
Data Source
AI summary
An arrangement for examining a plurality of samples within a sample region includes optics configured to selectively image the sample region in a transmitted light imaging mode or a fluorescence imaging mode. The arrangement further includes a processing apparatus configured to provide first operating instructions causing the arrangement to be operated in the transmitted light imaging mode, select one or more regions of interest in the sample region based on selection information obtained based on a stack of widefield images generated in the transmitted light imaging mode, provide position information relating to the one or more regions selected in the sample region, and provide second operating instructions causing the arrangement to be operated in the fluorescence imaging mode at fluorescence imaging positions based on the position information.