Fluorescence Observation Apparatus with Dynamic Exposure and Light Intensity Control
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Solution Overview
Problem
Conventional fluorescence observation apparatuses face challenges in achieving appropriate brightness and quantitativeness of fluorescence images due to variations in exposure time and light intensity, leading to potential over-excitation or under-excitation of fluorescent dyes, which affects the accuracy of fluorescence intensity correction.
Innovation Solution
The apparatus includes an illumination section with adjustable light sources for both illumination and excitation light, an exposure-time adjustment section, and normalization sections to control and normalize the luminance of fluorescence and reference images, ensuring constant brightness and accurate correction of fluorescence intensity changes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the exposure time is reduced to prevent over-excitation and fading of fluorescent dye, then the fluorescence image brightness is maintained, but the image may become too dark to observe
Solution Approach 1:
The system dynamically adjusts multiple parameters including exposure time, illumination light intensity, and excitation light intensity in a coordinated manner. By changing these parameters together rather than solely adjusting exposure time, the system maintains fluorescence image brightness while preventing over-excitation and dye fading.
Solution Approach 2:
The system uses the reference image luminance value as feedback to automatically adjust exposure time and light intensities. This closed-loop control ensures that the fluorescence image brightness is maintained at appropriate levels while preventing conditions that would cause dye fading or image saturation.
2Illumination intensity
If the intensity of excitation light is increased to improve fluorescence image brightness, then the fluorescence signal is enhanced, but the fluorescent dye fades and the image becomes dark
Solution Approach 1:
Instead of solely increasing excitation light intensity to improve brightness, the system coordinates multiple parameter adjustments including exposure time and illumination light intensity. This balanced approach enhances fluorescence signal while preventing dye fading through controlled exposure conditions.
Solution Approach 2:
The system monitors reference image luminance and uses this feedback to regulate excitation light intensity dynamically. This prevents excessive light intensity that would cause dye fading while maintaining sufficient brightness for observation.
3Illumination intensity
If the intensity of illumination light is changed to adjust image brightness, then the brightness is improved, but the corrected fluorescence image loses quantitativeness due to effects from simultaneous changes in excitation and illumination light intensities
Solution Approach 1:
The system adjusts illumination light intensity and excitation light intensity in a coordinated and controlled manner, along with exposure time. This multi-parameter coordination ensures that brightness adjustments do not compromise the quantitativeness of fluorescence intensity measurements, as all changes are systematically managed.
Solution Approach 2:
The system uses reference image luminance feedback to regulate both illumination and excitation light intensities together. This coordinated feedback control ensures that changes in one light source are compensated by corresponding adjustments in the other, maintaining measurement precision while adjusting brightness.
4Illumination intensity
If the exposure time is extended to improve image brightness, then the fluorescence image becomes brighter, but over-excitation occurs causing dye fading and image darkening
Solution Approach 1:
The system coordinates adjustments of exposure time with simultaneous adjustments of illumination light intensity and excitation light intensity. This prevents over-excitation by balancing the total light dose delivered to the sample, maintaining dye stability while achieving appropriate image brightness.
Solution Approach 2:
The system uses reference image luminance feedback to regulate exposure time dynamically. This prevents excessive exposure duration that would cause dye fading, while maintaining sufficient brightness through coordinated light intensity adjustments.
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 configuration allows for the acquisition of quantitative and appropriately bright fluorescence images by adjusting exposure time and light intensities, effectively correcting for changes in fluorescence intensity caused by distance variations, thereby enhancing the accuracy and quality of fluorescence observations.
Implementation Method 1
a light source for irradiating a subject with illumination light and excitation light
Implementation Method 2
acquires a fluorescence image from fluorescence produced in the subject irradiated with the excitation light
Implementation Method 3
acquires a reference image from return light returning from the subject irradiated with the illumination light
Data Source
AI summary
A quantitative fluorescence image and appropriate brightness is acquired and observed. Provided is a fluorescence observation apparatus including: an illumination section that includes a light source for irradiating an observation target region with illumination light and excitation light; a fluorescence image acquisition section that acquires a fluorescence image from fluorescence produced in the observation target region; a white-light image acquisition section that acquires a reference image from return light returning from the observation target region; an exposure-time adjustment unit that adjusts the exposure time based on the luminance value of the reference image; a diaphragm control section and a semiconductor laser control section that control the intensity of the illumination light and that of the excitation light based on the exposure time; a first normalization section that normalizes the luminance of the reference image and the fluorescence image by the exposure time; a second normalization section that normalizes the luminance of the reference image and the fluorescence image by the light intensity; andan image correction section that corrects the fluorescence image by the reference image, by using at least one of the normalized reference image or fluorescence image.