Fluorescence Microscope Imaging Control to Reduce Sample Bleaching
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Solution Overview
Problem
Current fluorescence microscopy techniques cause sample bleaching due to high illumination intensity and noisy, blurry live images, which are addressed by reducing illumination during still image streams when no image affecting changes are detected.
Innovation Solution
A control system switches between live and still image modes based on detected changes in microscope settings or sample location, using the last live image for still image streams and reducing illumination in still mode to minimize bleaching.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If high illumination intensity is used for live imaging, then sufficient fluorescence emission is achieved, but sample bleaching occurs
Solution Approach 1:
The illumination intensity is dynamically adjusted based on the imaging mode. During live imaging, high illumination intensity is applied to ensure sufficient fluorescence emission. During still image acquisition, the illumination intensity is reduced to minimize sample bleaching. This dynamic adjustment allows the system to optimize between image quality and sample preservation.
Solution Approach 2:
The system alternates between live imaging mode and still image mode in a periodic manner. Live imaging provides real-time observation with high illumination, while still image acquisition with reduced illumination preserves the sample. This periodic switching allows continuous monitoring while minimizing cumulative bleaching effects.
2Object-affected harmful factors
If low illumination power is used to minimize bleaching, then sample preservation is improved, but image quality becomes noisy and blurry
Solution Approach 1:
The system dynamically switches between two imaging modes: live imaging mode with high illumination intensity for sufficient signal quality, and still image mode with reduced illumination intensity for sample preservation. The control unit adjusts illumination power based on the selected mode, optimizing the trade-off between image quality and bleaching minimization.
Solution Approach 2:
During still image mode, the system uses a copy of the last live image as the display output while acquiring a new still image with reduced illumination. This allows the user to continue viewing the sample without interruption, while the reduced illumination protects the sample during the acquisition process.
3Illumination intensity
If high exposure time or high gain values are used, then live image visibility is improved, but image noise increases
Solution Approach 1:
The system dynamically adjusts camera parameters including exposure time and gain values based on the imaging mode. During live imaging, high exposure time and gain values are used to ensure sufficient image visibility. During still image acquisition, these parameters are optimized to reduce noise while maintaining image quality.
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
Significantly reduces bleaching effects and preserves the sample by optimizing illumination intensity, ensuring high-quality image presentation.
Implementation Method 1
the specimen is illuminated by a powerful light source with a certain wavelength. Such illumination causes the fluorescence dye to emit a certain emission wavelength at sufficient intensity
Data Source
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AI summary
The present invention relates to a microscope (120) including a control system (110) for controlling imaging of a sample (155) imaged by said microscope, wherein said microscope (120) comprises an illumination system (130) for illuminating said sample and an imaging system (140) for delivering microscopic images (170) of said sample, said control system (110) being connected to said illumination system (130) and to said imaging system (140) and being configured to detect an image affecting change in the microscope settings and/or in the location of the sample, which image affecting change results in an alteration of the sample image (170), and to cause the imaging system (140) to either deliver a live image stream (272) of live images of said sample in a first imaging mode, or to deliver a still image stream (274) of one or more still images of said sample in a second imaging mode, and to switch from the first imaging mode into the second imaging mode when no image affecting change is detected, using one of the last live images of the first imaging mode for at least a part of the still image stream (274) in the second imaging mode, and to reduce an illumination intensity of the illumination system (130) during the second imaging mode, and to a corresponding method. The invention especially aims at reducing bleaching of dyes and samples and protecting user eyes in fluorescence microscopy.