Inverted SPIM Microscopy for Multiwell Plate Imaging
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Solution Overview
Problem
Existing SPIM systems are not compatible with conventional specimen holders like multiwell plates or microfluidic chambers, and they lack the benefits of inverted microscopy such as easy access and quick specimen exchange without adjusting optics.
Innovation Solution
A SPIM system with an optical setup where the excitation and detection objectives are positioned beneath a stage, using an open-top, hollow prism to contact the specimen holder with liquid, allowing for imaging through glass coverslips or dipping objectives immersed in the liquid, enabling the use of conventional specimen holders and microfluidic devices.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If SPIM systems use conventional orthogonal objective placement, then volumetric imaging with reduced phototoxicity is achieved, but compatibility with conventional specimen holders and ease of operation are worsened
Solution Approach 1:
The patent inverts the conventional SPIM setup by placing objectives beneath the specimen holder rather than above it. This inversion enables compatibility with conventional specimen holders like multiwell plates and petri dishes while maintaining the light sheet imaging capability that reduces phototoxicity. The inverted configuration allows easy access to specimens from above without requiring optical adjustments.
2Measurement precision
If SPIM systems use orthogonal objective placement, then volumetric imaging is achieved, but adaptability to various specimen formats is worsened
Solution Approach 1:
The inverted SPIM configuration enables a single system to accommodate multiple specimen formats including multiwell plates, petri dishes, and microfluidic chambers. The objectives positioned beneath the stage can focus light sheets through the bottom of these diverse holders, providing universal compatibility while maintaining volumetric imaging capability.
3Reliability
If SPIM systems use conventional objective placement, then imaging capability is maintained, but time for specimen exchange is worsened
Solution Approach 1:
By inverting the objective placement to beneath the stage, the system enables quick specimen exchange from above without requiring optical realignment. Specimens can be rapidly replaced in conventional holders while the inverted objectives maintain consistent imaging geometry, eliminating time-consuming optical adjustments.
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 volumetric imaging with reduced phototoxicity and photobleaching, allowing for high-resolution imaging of specimens in various formats like petri dishes, multiwell plates, and microfluidic chambers, with improved accessibility and specimen exchange capabilities.
Implementation Method 1
a sheet of excitation light passes through the specimen and is coincident with the focal plane of a detection objective
Data Source
AI summary
In one embodiment, a selective plane illumination microscopy system for capturing light emitted by an illuminated specimen, the system including a specimen support having a top surface configured to support a specimen holder and an opening configured to provide access to a bottom of the specimen holder, and a selective plane illumination microscopy optical system positioned beneath the specimen support, the optical system configured to illuminate the specimen with a sheet of excitation light and including an excitation objective, a detection objective, and an open-top, hollow imaging element that is configured to contain a liquid, wherein the imaging element is positioned within the opening of the specimen support and optical axes of the objectives are aligned with the imaging element such that the axes pass through the imaging element and intersect at a position near the top surface of the specimen support.