Microscope Aberration Correction via Pre-calculated Holograms
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Solution Overview
Problem
In laser scanning microscopes, spherical aberrations occur due to mismatched refractive indices between biological samples and surrounding media, leading to weakened condensing intensity and spread of the condensing shape, especially when the sample surface is non-flat, such as in zebrafish or cell samples with varying refractive indices.
Innovation Solution
A microscope apparatus and method that includes a shape acquisition unit for surface and subsurface structure information, a hologram generation unit to create aberration correction data, and spatial light modulators to modulate the irradiation light, ensuring accurate aberration correction and maintaining condensing intensity and shape within the biological sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If aberration correction is applied using a spatial light modulator, then spot shape is improved, but device complexity increases due to additional components and control systems
Solution Approach 1:
The patent pre-calculates and stores aberration correction values for multiple Z-axis positions before actual imaging. The spatial light modulator applies these pre-computed corrections based on the current Z-position, eliminating the need for real-time complex calculations and reducing operational complexity while maintaining precise spot shape correction.
Solution Approach 2:
The system automatically selects and applies the appropriate aberration correction value based on the current Z-axis position without requiring manual intervention. The control unit autonomously manages the spatial light modulator settings, making the system self-adjusting and reducing operational complexity.
2Manufacturing precision
If high numerical aperture objective lens is used to improve resolution, then manufacturing precision is improved, but spherical aberration increases due to refractive index mismatch
Solution Approach 1:
The patent dynamically changes the wavefront parameters using a spatial light modulator to compensate for spherical aberration. By adjusting the phase parameters of the illumination light based on pre-calculated correction values for different Z-positions and numerical apertures, the system maintains high resolution while correcting spherical aberration caused by refractive index mismatch.
3Adaptability or versatility
If Z-axis position is changed to observe different depths, then adaptability is improved, but aberration correction accuracy decreases without position-specific calibration
Solution Approach 1:
The patent performs preliminary aberration correction measurements and stores correction values for multiple predetermined Z-axis positions. When the Z-position changes, the system automatically retrieves and applies the corresponding pre-calibrated correction value, maintaining high aberration correction accuracy across different depths without requiring real-time recalibration.
Solution Approach 2:
The system dynamically adapts the aberration correction by selecting different pre-calibrated correction values based on the current Z-axis position. This dynamic selection ensures that the appropriate correction is applied for each depth level, maintaining measurement precision while providing adaptability across different observation depths.
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 approach effectively suppresses the decrease in condensing intensity and spreading of the condensing shape, enhancing image clarity and resolution by correcting aberrations caused by surface and subsurface features of biological samples.
Implementation Method 1
controlling the wavefront of the irradiation light using a spatial light modulator and correcting the spherical aberration
Implementation Method 2
a spherical aberration is generated due to mismatch in refractive index between an observation target and a surrounding medium
Implementation Method 3
an objective lens; a light source for outputting light with which the biological sample is irradiated via the objective lens
Implementation Method 4
a condensing point of the irradiation light extends in an optical axis direction inside the observation target
Data Source
AI summary
A microscope apparatus (1A) includes a biological sample table (11) that supports the biological sample (B), an objective lens (12) disposed to face the biological sample table (11), a laser light source (13) that outputs light with which the biological sample (B) is irradiated via the objective lens (12), a shape measurement unit (20) that acquires a surface shape of the biological sample (B), a control unit (40) that generates aberration correction hologram data for correcting an aberration caused by the surface shape of the biological sample (B) on the basis of information acquired in the shape measurement unit (20), a first spatial light modulator (33) to which a hologram based on the aberration correction hologram data is presented and that modulates the light output from the laser light source (31), and a photodetector (37) that detects an intensity of light to be detected (L2) generated in the biological sample (B). Thus, a microscope apparatus and an image acquisition method capable of suppressing a decrease in condensing intensity of irradiation light inside a biological sample and spreading of a condensing shape are realized.