Microscope Autofocus via Laser Intensity Peak Detection
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Solution Overview
Problem
Conventional microscopy methods are time-consuming and inefficient for rapid and reliable focusing, especially in three-dimensional sample areas, particularly in fluorescence microscopy, due to the need for evaluating structural sharpness at multiple depth positions.
Innovation Solution
A method involving a laser beam directed onto a sample area with relative movement along the optical axis to detect intensity values, determining the highest intensity values, and using these to identify a reference relative travel position for precise focusing, allowing for rapid and reliable image capture without extensive filtering or processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods evaluate structural sharpness at multiple depth positions to determine optimal focus, then focusing accuracy is improved, but processing time increases significantly
Solution Approach 1:
The patent extracts only the most relevant information for focusing evaluation by detecting laser beam intensity at a specific point (focal point or focal plane) rather than evaluating entire images. This selective extraction of critical data enables rapid focusing determination without the time-consuming process of analyzing multiple depth positions or evaluating structural sharpness across the entire image field.
Solution Approach 2:
The patent replaces the mechanical/evaluative process of assessing structural sharpness with an optical measurement approach. Instead of capturing and analyzing multiple images at different depths to evaluate focus quality, the system uses a laser beam intensity measurement that directly indicates focus position, substituting complex image processing with a simpler optical detection method.
2Loss of information
If extensive patterns are projected onto the sample area for focusing evaluation, then focusing information is improved, but device complexity and processing requirements increase
Solution Approach 1:
The patent extracts only the essential focusing information by measuring laser beam intensity at a single point (the focal point or focal plane) rather than projecting and analyzing extensive patterns across the sample area. This approach obtains sufficient focusing information while avoiding the complexity of pattern projection systems and pixel-by-pixel evaluation.
Solution Approach 2:
Instead of projecting patterns onto the sample and evaluating reflections as done in prior art, the patent inverts the approach by directing the laser beam through the objective to focus onto the sample and detecting the intensity of the focused beam. This reverse approach simplifies the system by eliminating the need for complex pattern projection and reflection evaluation.
3Reliability
If multiple pixels within a virtual mask area are evaluated to derive focusing measure, then focusing reliability is improved, but processing time and computational load increase
Solution Approach 1:
The patent extracts the critical focusing information from a single point measurement (the focal point or focal plane) rather than evaluating multiple pixels within a virtual mask area. This extraction of the most relevant data point maintains focusing reliability by directly measuring where the beam focuses, while dramatically reducing processing time and computational requirements compared to merging intensity values from multiple pixels.
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
Enables quick and accurate focusing in microscopy, particularly in three-dimensional samples, by simplifying the evaluation process and reducing the need for precise filtering, thus enhancing throughput and reliability.
Implementation Method 1
detecting, for a respective relative travel position, a plurality of intensity values of the laser beam reflected at the interface and passing through the objective
Data Source
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AI summary
A method and a microscopy system for recording an image of a sample region are provided, including: directing a laser beam onto the sample region containing at least one interface by means of at least one objective lens, wherein the objective lens brings about an imaging of the laser beam on a focusing point which lies on the optical axis of the objective lens or on an axis lying parallel thereto and which further lies in a focusing plane; displacing the objective lens and the sample region with respect to one another in relative fashion along the optical axis of the objective lens to a plurality of different relative displacement positions; capturing a plurality of intensity values of the laser beam for a respective relative displacement position, said laser beam being reflected at the interface and passing through the objective lens, said intensity values being detected by pixels of a two-dimensional portion of a detection surface of a microscope camera; determining a respective highest intensity value for a respective displacement position; determining a curve of the highest intensity values; determining a reference relative displacement position from at least one maximum of the curve; capturing at least one image of the sample region at the reference relative displacement position.