Trans-illumination Imaging Meniscus Refraction Compensation
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Solution Overview
Problem
Trans-illumination imaging systems face challenges in achieving uniform contrast and image quality due to the detrimental effects of a liquid or semi-solid meniscus on the imaging of biological cells in multi-well sample holders, particularly at low magnification, where the meniscus disproportionately affects light rays and results in brighter central regions and darker edge regions.
Innovation Solution
The use of an array of independently energizable light sources that irradiate the sample with light beams at different orientations, allowing for differential energization to compensate for refraction by the meniscus, thereby improving contrast and uniformity across the image.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If a single light source or uniform illumination is used for trans-illumination imaging, then the imaging system is simple and easy to operate, but the image contrast is non-uniform with brighter central regions and darker edge regions due to meniscus refraction
Solution Approach 1:
The illumination system is divided into multiple independently controllable light sources arranged in an array, allowing different regions of the sample to be illuminated with different intensities. This segmentation enables compensation for the meniscus refraction effect by adjusting individual light source outputs to achieve uniform image contrast across the entire field of view.
Solution Approach 2:
Different light sources in the array are assigned different illumination intensities based on their position relative to the sample well. Light sources illuminating edge regions are intensified relative to those illuminating central regions, creating a non-uniform illumination pattern that compensates for the darkening effect of meniscus refraction at the edges.
2Reliability
If the meniscus is present in the well, then the sample can be contained and maintained, but refraction by the meniscus causes non-uniform light distribution and reduced image quality
Solution Approach 1:
The system pre-compensates for the known refraction effect of the meniscus by adjusting the illumination pattern before imaging. By calculating the expected refraction-induced intensity distribution and applying the opposite correction through differential light source energization, the system eliminates the harmful effect while maintaining the meniscus for sample containment.
Solution Approach 2:
The illumination parameters (intensity, distribution pattern) are dynamically adjusted based on the presence of the meniscus. The system modifies the energization levels of individual light sources in the array to counteract the refraction effect, transforming the uniform illumination parameter into a position-dependent illumination pattern that compensates for meniscus-induced non-uniformity.
3Area of stationary object
If low magnification imaging is used to capture the entire well, then the field of view is sufficient, but the contrast variation across different regions of the well floor is pronounced
Solution Approach 1:
The solution moves from considering illumination in a single dimension (uniform across the field) to a two-dimensional illumination pattern where intensity varies with position. By mapping light source intensities to spatial coordinates in the well, the system creates a gradient illumination pattern that compensates for the non-uniform contrast caused by low magnification imaging across the entire well area.
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 higher and more uniform contrast across the well floor, ensuring consistent image quality with different sample and well configurations, by adjusting the light contribution from each subset of light sources to equalize detected light intensity, thus enhancing the visibility of cells regardless of their position.
Implementation Method 1
compensate for refraction by the meniscus
Data Source
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AI summary
Method and system for trans-illumination imaging. In an exemplary method, a sample is irradiated in a well. The sample may include biological cells and a liquid or semi-solid medium that forms a meniscus. The step of irradiating may be performed with an array of light sources generating light of respective light beams that are incident on the meniscus at different orientations from one another. One or more images may be detected. A proportional contribution of light from two or more subsets of the light sources to the one or more images may be controlled by differential energization of the two or more subsets relative to one another to compensate for refraction by the meniscus and improve contrast.