Oblique Plane Microscopy With Single-Objective 90° Detection
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Solution Overview
Problem
Conventional optical microscopy techniques for obtaining optically sectioned images, such as confocal and structured illumination microscopy, face challenges including mechanical complexity, photodamage, phototoxicity, and noise accumulation, while Selective Plane Illumination Microscopy (SPIM) requires two objective lenses, limiting resolution and sample preparation flexibility.
Innovation Solution
An optical arrangement using a single high numerical aperture objective lens to illuminate and detect light at 90° angles, combined with a second optical subassembly to correct aberrations and produce an intermediate image with unity magnification, allowing for oblique plane imaging without mechanical complexity and conventional sample preparation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If confocal microscopy is used to obtain optically sectioned images, then image quality is improved, but data acquisition rate is reduced and photodamage increases
Solution Approach 1:
The patent replaces the mechanical scanning system of confocal microscopy with a stationary detection system. Instead of physically scanning a point detector across the sample, the invention uses a stationary detector that receives light from the entire field of view simultaneously, eliminating mechanical moving parts while maintaining optical sectioning capability through the specific optical arrangement with the 45-degree angled detector.
Solution Approach 2:
The patent transitions from one-dimensional point-by-point scanning to two-dimensional simultaneous detection by positioning the detector at a 45-degree angle to the optical axis, allowing the entire imaging plane to be detected at once rather than scanning through it sequentially.
2Productivity
If rapid scanning is employed to increase data acquisition rate, then productivity is improved, but peak power at the sample increases causing photodamage
Solution Approach 1:
The patent eliminates the need for rapid mechanical scanning by using a stationary detector configuration. The optical arrangement with the detector positioned at 45 degrees to the optical axis enables simultaneous detection of the entire field of view without mechanical movement, thereby avoiding the high peak powers associated with rapid scanning while maintaining high data acquisition rates.
3Measurement precision
If SPIM technique is used to illuminate a thin sheet, then optical sectioning is improved, but numerical aperture is reduced limiting resolution
Solution Approach 1:
The patent positions the detector at a 45-degree angle to the optical axis, creating a geometric arrangement where the detection plane is perpendicular to the illumination sheet. This angular configuration allows the use of high numerical aperture objectives without the mechanical constraints of SPIM, maintaining both thin optical sectioning and high resolution by decoupling the illumination and detection paths in three-dimensional space.
4Measurement precision
If two objective lenses are used in SPIM, then optical sectioning is improved, but device complexity increases
Solution Approach 1:
The patent combines the illumination and detection functions into a single objective lens system. Instead of using two separate objective lenses as in SPIM, the invention uses one objective for both illuminating the sample and collecting the emitted light, which is then directed to a stationary detector positioned at 45 degrees. This merging reduces mechanical complexity while maintaining optical sectioning capability.
Solution Approach 2:
The single objective lens in the patent serves multiple functions: it acts as both the illumination objective and the detection objective. This multi-functional design eliminates the need for two separate objectives and their associated mounting and alignment mechanisms, reducing device complexity while achieving the desired optical sectioning through the angular detector configuration.
5Measurement precision
If two objective lenses are used in SPIM, then optical sectioning is improved, but adaptability to sample preparation is reduced
Solution Approach 1:
The patent merges illumination and detection into a single objective path, allowing the use of standard glass microscope slides and conventional sample preparation techniques. The 45-degree angled detector configuration enables this single-objective approach to achieve optical sectioning without requiring the specialized sample holders and mounting arrangements needed for two-objective SPIM systems.
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
Enables high-resolution, low-aberration imaging of oblique planes using conventional sample preparations, reducing photodamage and noise, and facilitating 3D imaging without mechanical scanning or complex calculations.
Implementation Method 1
An optical arrangement uses a single high numerical aperture objective lens to illuminate and detect light at 90° angles
Implementation Method 2
combined with a second optical subassembly to correct aberrations and produce an intermediate image with unity magnification
Data Source
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AI summary
An optical arrangement for oblique plane microscopy comprising: a first optical subassembly, including an objective lens arranged to receive light from a sample in use, and configured to produce an intermediate image of the sample; and a second optical subassembly focused on the intermediate image, the optical axis of the second optical subassembly being at an angle to the optical axis of the first optical subassembly at the point of the intermediate image, such that the second optical subassembly images an oblique plane in the intermediate image, corresponding to an oblique plane in the sample. Also provided is a method of performing oblique plane microscopy comprising directing an incident beam of light through an objective lens to illuminate or excite an oblique plane in a sample, and receiving light from the sample through the same objective lens, wherein the incident beam of light is incident on the sample at an angle of substantially 90° relative to the beam of light received from the sample.