Microscope Activating Light Control for Fluorescent Density Bias
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Solution Overview
Problem
Conventional super-resolution microscopy methods struggle to obtain high-resolution images when there are significant variations in fluorescent material density within the same view field, as individual fluorescent bright spots cannot be separately detected in areas with high-density material.
Innovation Solution
A microscope apparatus and observation method that utilize total reflection illumination and acousto-optic tunable filters to control the intensity of activating light, allowing for selective activation and imaging of fluorescent material, thereby achieving uniform bright spot density across the view field.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If uniform activating light irradiation is applied across the view field, then simple operation is maintained, but high-resolution imaging fails in regions with high fluorescent material density due to overlapping bright spots
Solution Approach 1:
The patent applies local quality by varying the irradiation intensity of activating light across different regions of the view field according to the fluorescent material density distribution. Regions with high fluorescent material density receive lower irradiation intensity, while regions with low density receive higher intensity, ensuring uniform bright spot density across the entire field of view and enabling high-resolution imaging in all areas
Solution Approach 2:
The patent implements preliminary action by calculating the two-dimensional intensity map based on preliminary imaging data before the main high-resolution imaging process. This preliminary map predicts the fluorescent material density distribution and pre-configures the optimal irradiation intensity distribution, allowing the system to adapt to different samples without real-time complex adjustments during image acquisition
2Quantity of substance
If high irradiation intensity of activating light is used, then bright spots are easily detected, but fluorescent material in high-density regions becomes saturated and bright spots overlap
Solution Approach 1:
The patent applies local quality by spatially varying the irradiation intensity of activating light across the view field. Regions with high fluorescent material density are irradiated with lower intensity to prevent saturation and maintain bright spot separation, while regions with low density receive higher intensity to ensure sufficient signal. This localized intensity control enables uniform bright spot density across the entire field of view
Solution Approach 2:
The patent implements parameter changes by dynamically adjusting the irradiation intensity parameter of activating light based on the calculated two-dimensional intensity map. The system modifies the light intensity parameter in response to the spatial distribution of fluorescent material, transforming a fixed-parameter approach into a variable-parameter approach that adapts to local conditions
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 the acquisition of high-resolution images even in cases of large bias in fluorescent material distribution by adjusting the irradiation intensity of activating light based on the two-dimensional intensity map, ensuring proper separation and detection of fluorescent bright spots.
Implementation Method 1
utilize total reflection illumination and acousto-optic tunable filters to control the intensity of activating light
Implementation Method 2
utilize total reflection illumination to control the intensity of activating light
Implementation Method 3
fluorescent material which is activated when irradiated with activating light of a prescribed wavelength, and which has the property of becoming inactive by emitting fluorescence when subsequently irradiated with exciting light
Data Source
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AI summary
A microscope apparatus includes: a distribution measurement apparatus which - with respect to an observation region wherein a sample is arranged which comprises a fluorescent material that is activated when irradiated with an activating light of a prescribed wavelength, and that emits fluorescence when irradiated in an activated condition with an exciting light of a wavelength that differs from that of the activating light - obtains a fluorescent picture image by conducting irradiation with the exciting light, and measures a fluorescent intensity distribution of the observation region; an irradiation intensity setting apparatus which sets irradiation intensities of the activating light for respective portions of the observation region based on the fluorescent intensity distribution; and a picture image formation apparatus which obtains a plurality of the fluorescent picture images by multiply repeating operations comprising an operation wherein the observation region is irradiated with the activating light at the irradiation intensities that have been set in the respective portions, and an operation wherein a fluorescent picture image is obtained by irradiating the observation region with the exciting light after the activating light irradiation, and which generates a sample picture image from the plurality of the fluorescent picture images.