Dynamic Correction Factor for Confocal Microscopy Aberrations

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Solution Overview

Problem

Current microscopic imaging methods face challenges in achieving high-resolution three-dimensional representation of specimens with low illumination intensities while ensuring high detection radiation efficiency and real-time imaging, often resulting in imaging aberrations and incorrect surface calculations due to specimen-independent correction factors.

Innovation Solution

A microscopic imaging method that dynamically determines a specimen-specific correction factor for each imaging session by capturing wide-field and composite signals, allowing for accurate extraction of confocal images and reducing aberrations by adjusting the correction factor based on the specimen's unique intensity ratios and characteristics.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If a specimen-independent correction factor is used for confocal image extraction, then the imaging process is simplified and faster, but imaging aberrations occur and measurement precision deteriorates

Engineering Contradiction:

Improveimaging speedVSAvoidsurface calculation accuracy

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The correction factor is transformed from a static, pre-determined value to a dynamic parameter that is recalculated for each imaging session. The system automatically determines the correction factor based on actual wide-field and composite image signals captured during each measurement, adapting to specimen-specific characteristics and eliminating the need for manual calibration while maintaining high imaging speed.

Inventive Principle:

Principle #15Dynamics

Solution Approach 2:

The imaging system performs self-calibration by automatically determining the correction factor using the captured wide-field and composite image signals. The system uses its own measurement data to calculate the appropriate correction factor without requiring external calibration standards or manual intervention, enabling both high speed and high precision.

Inventive Principle:

Principle #25Self-service

2Device complexity

If a fixed correction factor is applied to all specimens, then device complexity is reduced, but adaptability to different specimen types deteriorates

Engineering Contradiction:

Improvecorrection factor calibrationVSAvoidspecimen-specific imaging accuracy

Core Design Contradiction:

Device complexityVSAdaptability or versatility

Solution Approach 1:

The correction factor parameter is changed dynamically based on the specific specimen being imaged. Instead of using a fixed parameter value, the system calculates the correction factor for each imaging session based on the actual optical paths and specimen characteristics, enabling the system to adapt to different specimen types including strongly reflecting and autofluorescing materials without increasing device complexity.

Inventive Principle:

Principle #35Parameter changes

3Object-affected harmful factors

If low illumination intensities are used to protect the specimen, then specimen damage is reduced, but detection signal strength decreases

Engineering Contradiction:

Improvespecimen damage from illuminationVSAvoiddetection radiation intensity

Core Design Contradiction:

Object-affected harmful factorsVSUse of energy by moving object

Solution Approach 1:

The system uses feedback from the captured wide-field and composite image signals to optimize the correction factor and enhance the confocal signal extraction. By applying the dynamically determined correction factor, the system maximizes the usable signal from low-intensity illumination, enabling high-quality imaging while minimizing specimen exposure to damaging radiation intensities.

Inventive Principle:

Principle #23Feedback

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 enhances signal quality, reduces imaging aberrations, and enables the examination of various materials, including strongly reflecting and autofluorescing specimens, by adapting the correction factor for each measurement, thereby improving the accuracy and reliability of three-dimensional imaging.

Implementation Method 1

capturing detection radiation along a detection axis, said detection radiation having been caused by the illumination radiation

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Data Source

PatentUS11238575B2Microscopic imaging method using a correction factor

Publication Date: 2022.02.01 CARL ZEISS MICROSCOPY GMBH

AI summary

A microscopic imaging method, includes illuminating a specimen with illumination radiation and capturing detection radiation along a detection axis. The detection radiation is caused by the illumination radiation, at a first time as a wide-field signal and at a second time as a composite signal. The composite signal is formed by a superposition of a confocal image and a wide-field image; extracting the confocal image by subtracting the wide-field signal from the composite signal, wherein a correction factor is used. A current correction factor is ascertained for each executed imaging and/or for each imaged specimen (1) and the confocal image is extracted using the respective current correction factor.