Microlens Wavefront Analysis for Wide-Field Fluorescence Microscopy
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Solution Overview
Problem
Existing microscopic imaging methods face challenges in achieving high-resolution and contrast imaging of complex biological objects due to optical defects and inhomogeneous refractive indices, particularly when imaging deep within the object, and current wavefront analysis methods require complex object preparation and are limited by the isoplanetary domain, which restricts the field of view.
Innovation Solution
A wavefront analysis device using a microlens array with a field diaphragm to measure local wavefront gradients and deviations, eliminating the need for artificial star generation, and allowing accurate wavefront analysis over larger fields by controlling image size and minimizing overlaps between microlens images, while using structured illumination for improved correlation calculations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Area of stationary object
If a wavefront analyzer without field diaphragm is used, then the field of view is larger, but image overlap between microlenses occurs causing reduced measurement precision
Solution Approach 1:
The patent divides the field of view into multiple discrete regions, each imaged by a separate microlens. The field diaphragm creates distinct image zones that prevent overlap between adjacent microlens images, allowing each microlens to independently measure wavefront characteristics in its designated region without interference from neighboring regions.
Solution Approach 2:
The field diaphragm introduces spatially varying optical properties across the field of view. By controlling the size and position of the field diaphragm, the patent optimizes the image size and separation for each local region, ensuring that images from different microlenses are properly separated while maximizing the usable field of view for wavefront analysis.
2Area of stationary object
If the field diaphragm size is increased, then the analyzed field is larger, but image overlap between microlenses increases
Solution Approach 1:
The field diaphragm is designed with variable size and position, allowing dynamic adjustment of the analyzed field. This enables the system to adapt the field diaphragm dimensions to match the specific requirements of different microlens arrangements and imaging conditions, optimizing the balance between field size and image separation for each measurement scenario.
3Measurement precision
If direct wavefront measurement methods are used, then measurement accuracy is improved, but device complexity and object preparation requirements increase
Solution Approach 1:
The patent uses the fluorescence image of the object itself as a reference copy for wavefront measurement. Instead of requiring external point sources or complex artificial stars, the system captures the fluorescence emission from the object and uses its intensity distribution as the reference pattern for correlation-based wavefront gradient measurement, significantly simplifying the instrumentation.
Solution Approach 2:
The object's own fluorescence emission serves dual purposes: it provides both the measurement signal and the reference pattern needed for wavefront analysis. The system uses the object's intrinsic properties (fluorescence distribution) to perform self-characterization, eliminating the need for external calibration sources or additional reference objects.
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 proposed method provides significantly improved accuracy in wavefront analysis and image correction, enabling high-resolution imaging over larger isoplanetary domains without the need for artificial star generation, suitable for complex biological structures like neural networks in neuroimaging.
Implementation Method 1
a microlens array configured for forming, in a detection plane, images of an optical section of the object
Implementation Method 2
a field diaphragm arranged in an intermediate focal plane of the microlens array, said field diaphragm being configured for limiting a size of the imaged field for each of said microlenses
Implementation Method 3
an illumination channel for illuminating an optical section of a volumetric and fluorescent object; a microscope objective including a pupil in a pupillary plane for receiving the fluorescence emission light from the object
Data Source
Figure 1
Figure 2A~2B
Figure 3
AI summary
According to one aspect, the present description relates to a device (110) for analysing a wavefront, configured to be connected to a fluorescence microscopy imaging system (100) with optical sectioning, equipped with a microscope objective (130) comprising a pupil in a pupil plane (P4), the analysis device comprising a two-dimensional detector (112) comprising a detection plane (P3); a two-dimensional arrangement (114) of microlenses (115), arranged in an analysis plane (P5), each microlens being configured to form, on the detection plane, when the analysis device is connected to the microscopic imaging system, an image of an object situated in a focal plane (P1) of the microscope objective, with a given analysis field; an optical relay system (116) configured to optically conjugate the analysis plane and the pupil plane (P4); a field diaphragm (118) positioned in a plane (P2) optically conjugated with the plane of detection (P3), and configured to define said analysis field; a processing unit (120) configured to determine, based on the set of images formed by the microlenses, a two-dimensional map of a characteristic parameter of the wavefront in said analysis plane.