Fluorescence Observation Apparatus Spatial Multiplexing
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Solution Overview
Problem
Current fluorescence observation methods experience increased image-capturing time as the number of observation-target dyes increases, due to the need for multiple filter switches and time-division wavelength adjustments.
Innovation Solution
A fluorescence observation apparatus and method using a stage, excitation section, and spectroscopic imaging section, where multiple line illuminations of different wavelengths are applied parallel to each other, allowing separate fluorescence reception and spatial separation of excitation wavelengths, reducing the need for temporal switching and thus minimizing image-capturing time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple filter switches are used to capture different dye spectra, then color separation is achieved, but image-capturing time increases linearly with the number of colors
Solution Approach 1:
The patent transitions from temporal separation (time-division) to spatial separation by arranging multiple bandpass filters in parallel along the optical path. Each filter captures a specific dye spectrum simultaneously, converting the time-based sequential capture into a spatial parallel capture system, thereby eliminating the linear increase in capture time.
Solution Approach 2:
The optical system is segmented into multiple parallel detection channels, each equipped with a dedicated bandpass filter for specific dyes. This segmentation allows simultaneous capture of multiple dye spectra through different spatial paths, resolving the contradiction between color separation precision and capture time.
2Measurement precision
If excitation light wavelength is switched to excite different dyes, then selective dye excitation is achieved, but image-capturing time increases with the number of wavelengths to be switched
Solution Approach 1:
The patent introduces a spatial dimension to the excitation system by arranging multiple line illumination sources at different positions along the optical axis, each emitting at a specific excitation wavelength. This allows simultaneous excitation of multiple dyes through spatially separated illumination paths, eliminating the need for temporal wavelength switching.
Solution Approach 2:
The excitation system is divided into multiple independent line illumination channels, each targeting specific dyes. This segmentation enables parallel excitation of multiple fluorophores simultaneously, resolving the time penalty associated with sequential wavelength switching while maintaining excitation selectivity.
3Measurement precision
If fluorescence from adjacent dyes leaks through bandpass filters, then color mixture occurs, but using narrower filters increases the number of filters needed
Solution Approach 1:
The detection system is segmented into multiple specialized channels, each with a bandpass filter optimized for a specific dye's emission spectrum. This dedicated channel assignment ensures that each detector receives primarily the signal from its target dye, minimizing cross-contamination and color mixture while maintaining a manageable filter count through efficient spectral matching.
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 prevents the increase in image-capturing time associated with multiple dyes, enabling efficient multiplexed imaging by spatially separating excitation wavelengths, thereby maintaining or reducing image acquisition time even with increased dye numbers.
Implementation Method 1
an excitation section that irradiates a pathological specimen on a stage with a plurality of line illuminations of different wavelengths
Implementation Method 2
a spectroscopic imaging section that includes at least one imaging device capable of separately receiving pieces of fluorescence respectively excited with the plurality of line illuminations
Data Source
AI summary
A fluorescence observation apparatus according to an embodiment of the present technology includes a stage, an excitation section, and a spectroscopic imaging section. The stage is capable of supporting a fluorescently stained pathological specimen. The excitation section irradiates the pathological specimen on the stage with a plurality of line illuminations of different wavelengths, the plurality of line illuminations being a plurality of line illuminations situated on different axes and parallel to a certain-axis direction. The spectroscopic imaging section includes at least one imaging device capable of separately receiving pieces of fluorescence respectively excited with the plurality of line illuminations.