Bright-field Reflection Microscope Annular Illumination
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Solution Overview
Problem
Bright-field reflection microscopes experience spurious resolution in three-dimensional image formation due to phase shifts in reflected light from the cover glass when the sample is moved along the optical axis, causing interference with the sample's reflected light and leading to incorrect object image representation.
Innovation Solution
The implementation of a bright-field reflection microscope with an illumination optical system using a plurality of annulus illuminations with different annulus radii, which are sequentially switched to generate annulus illumination patterns, and a processing unit that processes capturing results to generate a three-dimensional image by compensating for phase shifts and interference effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If typical illumination such as Köhler illumination is used to illuminate the sample, then the sample can be observed with reflected light, but spurious resolution occurs in the three-dimensional image due to phase shifts in reflected light from the cover glass
Solution Approach 1:
The illumination optical system divides the illumination light into multiple annular beams with different annulus radii using an aperture pattern turret. Each annular beam has a specific radius that corresponds to a particular imaging depth, allowing selective illumination of different focal planes. This segmentation of the illumination beam enables the system to capture multiple three-dimensional images at different depths without spurious resolution, as each annular beam illuminates a specific focal plane independently.
Solution Approach 2:
The patent introduces a new dimension of control by varying the annulus radius of the illumination beams. Instead of using a single uniform illumination beam, the system creates multiple annular beams with different radii, adding a radial dimension to the illumination pattern. This dimensional change allows the system to selectively illuminate different focal planes and captures three-dimensional information without the phase shift artifacts that occur with conventional illumination methods.
2Measurement precision
If the sample is driven in the optical axis direction to capture three-dimensional images, then depth information can be obtained, but the phase of reflected light from the cover glass shifts causing incorrect object image representation
Solution Approach 1:
The illumination system segments the illumination light into multiple annular beams with different radii, where each annular beam is designed to illuminate a specific focal plane. When the sample is driven in the optical axis direction, each annular beam maintains its phase relationship with the corresponding focal plane, allowing accurate capture of three-dimensional images without phase shift artifacts. The aperture pattern turret selectively positions different annular aperture patterns to match the focal plane being imaged.
Solution Approach 2:
The annular aperture patterns act as intermediaries between the illumination light and the sample. These aperture patterns selectively transmit light at specific annulus radii that correspond to particular imaging depths. By using these intermediary aperture patterns, the system can control which focal plane is illuminated and captured, preventing phase shift errors from propagating to the final image while still enabling three-dimensional imaging through optical axis movement.
3Device complexity
If a single illumination beam is used to illuminate the sample, then the optical system is simple, but spurious resolution occurs due to interference between reflected light from the sample and cover glass
Solution Approach 1:
The illumination optical system uses an aperture pattern turret to segment the illumination beam into multiple annular beams with different radii. Each annular beam is designed to illuminate a specific focal plane, and the aperture pattern turret can selectively position different annular aperture patterns. This segmentation increases the device complexity by adding the aperture pattern turret and multiple aperture patterns, but it eliminates spurious resolution by ensuring that each annular beam illuminates only its corresponding focal plane without interfering with other planes.
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 effectively reduces spurious resolution, allowing for accurate three-dimensional image reconstruction of samples by compensating for phase shifts and interference, resulting in a more precise representation of the sample's structure.
Implementation Method 1
an illumination optical system that generates each of a plurality of annular illumination lights having annulus radiuses different from one another and illuminates the sample with the illumination lights
Implementation Method 2
gathers a first reflected light from the sample and a second reflected light from an interface of surroundings of the sample
Implementation Method 3
gathers... at the capturing device via the objective lens
Implementation Method 4
detects the first reflected light and the second reflected light at each of the plurality of positions
Implementation Method 5
generating an image frequency in a frequency space from a plurality of capturing results obtained at each of the plurality of positions... by using each of the plurality of annular illumination lights
Data Source
AI summary
A bright-field reflection microscope according to the present embodiment includes an illumination optical system that includes an aperture pattern turret that can form a plurality of annular illumination lights having annulus radiuses different from each other and an objective lens and illuminates the sample S with the illumination light; a detection optical system that gathers a first reflected light from the sample S and a second reflected light from an interface of surroundings of the sample S at the capturing device via the objective lens; and a control unit, and the capturing device detects the first reflected light and the second reflected light at each of the plurality of positions with different relative positions to the objective lens and the sample S by using each of the plurality of annular illumination lights formed by the control unit controlling the aperture pattern turret.