Fluorescence Microscope Threshold Adjustment for Noise Reduction

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Solution Overview

Problem

Existing fluorescence microscopy methods face challenges in selecting a suitable evaluation parameter for generating high-quality images of structures or processes within a sample, often resulting in noise-induced light spots being misclassified as events.

Innovation Solution

A method where a signal representative of the pictorial light distribution is generated, allowing for user-adjustable threshold values to mark and classify areas with comparison values greater than the threshold, thereby optimizing the selection of evaluation parameters and reducing noise classification.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If a low threshold value is used to classify light spots as events, then more fluorescent dye particle events are detected, but noise-induced light spots are misclassified as events

Engineering Contradiction:
Improveevent detection sensitivityVSAvoidclassification accuracy
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

The system displays the pictorial light distribution with marked sub-areas to the user, who can then adjust the threshold value based on visual feedback. This iterative feedback loop allows the user to optimize the threshold setting to achieve both high sensitivity and accuracy in event classification.

Inventive Principle:
Principle #23Feedback

Solution Approach 2:

The user directly adjusts the threshold parameter based on their knowledge of the sample and visual inspection of the displayed distribution. This self-service approach allows expert users to leverage their domain knowledge to set optimal thresholds for specific experimental conditions.

Inventive Principle:
Principle #25Self-service

2Reliability

If a high threshold value is used to filter out noise, then classification accuracy improves, but genuine fluorescent dye particle events are missed

Engineering Contradiction:
Improveclassification accuracyVSAvoidevent detection sensitivity
Core Design Contradiction:
ReliabilityVSMeasurement precision

Solution Approach 1:

The visual display of marked sub-areas provides immediate feedback to the user about which regions are being classified as events. This allows the user to adjust the threshold to ensure genuine events are not missed while still filtering noise effectively.

Inventive Principle:
Principle #23Feedback

Solution Approach 2:

The threshold value is made adjustable by the user, allowing dynamic optimization of the classification parameter based on specific experimental conditions, sample characteristics, and noise levels observed in the data.

Inventive Principle:
Principle #35Parameter changes

3Reliability

If manual threshold optimization is performed, then classification accuracy improves, but time consumption increases

Engineering Contradiction:
Improveevaluation parameter suitabilityVSAvoidparameter setting time
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The system provides visual feedback of the current threshold setting through the display of marked sub-areas, enabling rapid assessment and adjustment. This reduces the time needed for manual optimization compared to trial-and-error approaches without visual feedback.

Inventive Principle:
Principle #23Feedback

Solution Approach 2:

The system creates a visual copy or representation of the light distribution with marked regions, allowing the user to assess the threshold setting quickly without having to manually analyze raw data or perform time-consuming calculations.

Inventive Principle:
Principle #26Copying

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 enables optimal selection of threshold values, ensuring that only genuine fluorescent dye particle events are classified, thereby improving image quality and reducing noise-induced errors.

Implementation Method 1

Fluorescent dye particles in a sample are excited to fluoresce, and the fluorescence light emitted by the dye particles is detected

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

The fluorescence emitted by a subset of active dye particles is imaged onto a spatially resolved light detector, for example, a CCD camera

Methodology Applied
Scientific EffectLight transmission and imaging: Light

Data Source

PatentEP2534521B1Method and device for setting a suitable evaluation parameter for a fluorescence microscope
Publication Date: 2019.03.27 LEICA MICROSYSTEMS CMS GMBH
  • EP2534521B1 patent drawingFigure 1~5
  • EP2534521B1 patent drawingFigure 6~7

AI summary

In order to set a suitable evaluation parameter for a fluorescence microscope (20), fluorescent dye particles are excited in a sample (32) so as to fluoresce. Fluorescent light originating from the dye particles is detected. A graphical light distribution is determined, which is representative of a distribution of the light quantity of the fluorescent light. A signal is generated, which is representative of the graphical light distribution and depending on which a display unit (44) is actuated such that the display unit (44) displays the graphical light distribution. A comparison value is associated with portiosn of the graphical light distribution, said value being representative of the light quantity in the corresponding portion. A threshold value is predefined as an evaluation parameter. The comparison values are compared to a predefined threshold value. The portions are marked on the display unit (44) with predefined markings (52), the comparison value of which is greater than the threshold value. Depending on a user input, the threshold value is changed. After the user input, the comparison values are compared to the changed threshold value. The marked portions are defined as events (53). An overall image of the sample (32) is determined depending on the events (53).