Cell Sample Imaging Using Contrast-Based Depth Focusing
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Solution Overview
Problem
Existing autofocus systems for microscopes struggle to accurately determine the optimal focus depth for cell samples, particularly in three-dimensional imaging, leading to inefficiencies in image capture and analysis.
Innovation Solution
A method and system for determining a reference depth level within a cell sample by analyzing image contrast through a series of depth scans, identifying a drop in contrast to establish the reference depth level, and using this information to focus the microscope for optimal imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If classical autofocus methods using standard deviation, entropy, or gradient functions are used to determine focus depth, then the focus function can be computed to identify the maximum sharpness or contrast level, but the method fails to accurately determine the reference depth level in three-dimensional cell samples, leading to incorrect focus positioning
Solution Approach 1:
Instead of searching for the maximum contrast or sharpness value to determine focus depth, the patent inverts the approach by detecting the minimum contrast value (the well) in the contrast curve. This inversion allows the system to identify the reference depth level corresponding to the bottom of the well, which provides accurate focus positioning in three-dimensional cell samples.
2Manufacturing precision
If depth scanning is performed to capture multiple images at different focal planes, then the microscope can capture images throughout the sample depth, but the process is time-consuming and reduces imaging efficiency
Solution Approach 1:
The patent performs a preliminary depth scanning to capture images at multiple focal planes and construct a contrast curve, but then uses this pre-acquired data to automatically determine the reference depth level without requiring additional manual intervention. This preliminary action enables rapid and accurate focusing in subsequent imaging operations.
Solution Approach 2:
The system uses the contrast curve generated during depth scanning as feedback to automatically identify the reference depth level. By continuously monitoring contrast changes during the scanning process and using this feedback to determine the minimum contrast point, the system achieves both accurate focusing and efficient image capture.
3Ease of operation
If manual focus adjustment is used to position the investigation zone on the optical axis and superimpose it with the focus plane, then the user can control the imaging parameters, but the process requires complex three-dimensional adjustment and reduces operational efficiency
Solution Approach 1:
The patent implements an automated autofocus system that performs depth scanning and contrast analysis without requiring manual user intervention. The system self-determines the reference depth level by detecting the minimum contrast value in the contrast curve, eliminating the need for manual three-dimensional adjustment and significantly reducing focus positioning time.
Data Source
AI summary
Apparatus and methods are described for use with a cell sample that includes a plurality of cells. A series of images associated with a series of depth levels of the cell sample are acquired, by performing a depth scan of cell sample with a microscope. One of the depth levels is identified as being an optimum focal plane for imaging one or more entities within the cell sample using the microscope, under a first illumination condition. The cell sample is imaged under a second illumination condition that is different from the first illumination condition, using the microscope, by focusing the microscope at an investigative depth level that is based on the identified depth level. Other applications are also described.