Bright Field Cell Imaging via Focus Difference Processing
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Solution Overview
Problem
Conventional techniques for imaging transparent cells are complex and costly, requiring phase difference microscopes with narrow imaging fields and high numerical aperture lenses, and often necessitate determining the in-focus position, which complicates the process and apparatus configuration.
Innovation Solution
An image processing method involving a multi-stage adjustment of the focus position in the optical axis direction to capture multiple images, calculating contrast values, and generating a difference image between images taken at positions on either side of a local minimum contrast value, allowing for clear contour visualization without needing to determine the in-focus position or using a phase difference imaging setup.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a phase difference microscope is used to image transparent cells, then image contrast is improved, but apparatus complexity and cost increase
Solution Approach 1:
The patent creates a virtual phase difference image by computationally processing two bright field images taken at different focus positions. Instead of using a physical phase difference microscope, the method copies the effect of phase difference imaging through image processing, where the difference between two defocused images simulates the phase contrast effect, thereby achieving high contrast imaging with a simple bright field microscope
Solution Approach 2:
The patent replaces the mechanical/optical complexity of a phase difference microscope with a computational approach. By substituting the physical phase difference optical system with digital image processing operations on bright field images, the method achieves the same imaging goal without requiring specialized optical components, thus reducing apparatus complexity and cost
2Measurement precision
If in-focus position determination is performed to obtain high quality images, then image quality is improved, but processing complexity increases
Solution Approach 1:
The patent inverts the conventional approach by not seeking the in-focus position for optimal imaging, but rather intentionally using two defocused positions to generate the final image. Instead of adjusting focus to maximize image quality directly, the method uses defocused images and processes their difference to achieve enhanced contrast, thereby eliminating the need for complex focus determination procedures
3Measurement precision
If phase difference imaging is used to obtain clear cell contours, then contour visibility is improved, but imaging field of view is reduced
Solution Approach 1:
The patent makes the bright field microscope multi-functional by enabling it to perform both standard bright field imaging and virtual phase difference imaging through software processing. The same optical system can capture images in conventional mode and then process them computationally to achieve phase difference effects, thereby maintaining a wide imaging field of view while obtaining clear cell contours without requiring specialized phase difference optical components that would limit the field of view
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 simplifies the apparatus configuration and image processing, enabling the generation of images with clear cell contours using a general bright field imaging setup, eliminating the need for phase difference imaging and in-focus position determination.
Implementation Method 1
imaging the cells in a bright field
Data Source
Figure 1
Figure 2A~2B
Figure 3(a)~3(c)
AI summary
An image processing apparatus comprises a holder 11 which holds a carrier WP carrying cells, an imager 13 which includes an imaging optical system 130 arranged such that an optical axis OA is oriented toward the carrier WP and images the cells in a bright field by receiving light incident on the imaging optical system, a position changer 146 which changes a focus position of the imaging optical system in a direction along the optical axis OA, a controller 141 which changes a set position of the focus position by the position changer in a multi-stage manner and obtaining a plurality of original images by causing the imager to perform imaging at each set position, and an image processor 141 which calculates a contrast value of each original image imaged at each set position, specifying two set positions at opposite sides of a local minimum value of the contrast value in a profile of the contrast value in relation to the set position and generating a difference image of two of the original images respectively imaged at the two set positions.