Light-Sheet Microscope Radiation Width Control
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Solution Overview
Problem
Microscopes using the light-sheet illumination technique face challenges in preventing unwanted exposure of specimens to excitation light, especially when magnification is switched, and in acquiring clear images over a wide area with insufficient depth of focus, particularly when excitation light beams of the same or overlapping wavelengths are used.
Innovation Solution
A microscope with a detection optical system and a light-sheet illumination optical system that controls the radiation width of excitation light and processes images by combining fluorescence images acquired at different focal positions, allowing for clear fluorescence imaging even with overlapping wavelengths, and alternately switching the incidence direction of excitation light to prevent mixing and enhance focus.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the magnification of the detection optical system is increased, then the imaging resolution is improved, but the radiation width of excitation light becomes too large causing unwanted exposure of the specimen
Solution Approach 1:
The patent applies local quality by adjusting the radiation width of excitation light to match the specific field of view at each magnification level. The control section dynamically modifies the illumination parameters locally to the observed region, ensuring that excitation light is confined only to the area being imaged at high magnification, thereby preventing unwanted exposure while maintaining imaging resolution.
2Measurement precision
If excitation light beams with the same wavelength are radiated on overlapping regions to acquire clear fluorescence images, then the image clarity is improved, but the time required for image acquisition increases due to sequential scanning
Solution Approach 1:
The patent employs periodic action by alternately switching between two different incidence directions of excitation light beams. The control section sequentially radiates excitation light from the first direction, then switches to the second direction, and repeats this periodic switching. This allows overlapping regions to be excited from multiple angles in an efficient sequence, maintaining image clarity while reducing total acquisition time compared to traditional sequential scanning from a single direction.
3Area of stationary object
If the depth of focus is increased to capture wide area images, then the coverage area is improved, but the image sharpness deteriorates due to insufficient depth of focus
Solution Approach 1:
The patent applies dimensionality change by introducing a second incidence direction for excitation light in addition to the first direction. By radiating excitation light from two different angular dimensions, the system effectively extends the depth of focus coverage. The control section coordinates excitation from both directions to capture fluorescence from different depth planes, thereby achieving sharp images across a wider area without compromising image sharpness.
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
The solution enables clear fluorescence imaging over a wide area with reduced unwanted exposure and improved depth of focus, allowing for high-speed image acquisition even with overlapping wavelengths, and reduces the time required for image acquisition by optimizing the incidence direction of excitation light.
Implementation Method 1
a detection optical system that detects fluorescence emitted from a specimen
Implementation Method 2
detects fluorescence emitted from a specimen
Implementation Method 3
a light-sheet illumination optical system that causes planar excitation light to be incident along a direction intersecting an optical axis of the detection optical system
Data Source
AI summary
To prevent unwanted exposure of a specimen with excitation light despite magnification switching, a microscope of the present invention includes: a detection optical system that detects fluorescence emitted from the specimen; a light-sheet illumination optical system, serving as an excitation light source, that causes planar excitation light to be incident along a direction intersecting an optical axis of the detection optical system; and a control section that, when the magnification of the detection optical system is raised, controls the light-sheet illumination optical system so as to reduce a radiation width of the excitation light made incident on the specimen by the light-sheet illumination optical system.