Folded Oblique Plane Microscopy for Compact 3D Scanning
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Solution Overview
Problem
Existing oblique plane microscopy (OPM) configurations require a long physical optical path and complex sample preparation due to the need for multiple high numerical aperture microscope objectives placed at significant angles to each other, making it difficult to achieve a compact and efficient scanning mechanism for 3D imaging.
Innovation Solution
A 'folded' OPM configuration that uses a first mirror located at the focal plane of a high numerical aperture objective lens to reflect both the illumination beam and emitted light, combined with a beam splitter to direct these beams efficiently, allowing for a more compact arrangement and enabling scanning without adjusting the axial position of remote objectives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple high numerical aperture microscope objectives are placed at significant angles to each other, then oblique plane microscopy can be achieved, but the physical optical path becomes long and the arrangement becomes complex
Solution Approach 1:
The patent introduces a folded optical path configuration that redirects the light path in multiple dimensions using mirrors and beam splitters. Instead of a straight linear path, the illumination and detection beams are folded back on themselves, effectively reducing the physical footprint while maintaining the required optical path length and angular relationships between objectives
Solution Approach 2:
The patent implements a nested optical arrangement where the detection optical path is positioned within or alongside the illumination optical path. The beam splitter and mirrors are arranged so that detection beams travel through portions of the same optical space as illumination beams, allowing compact integration of multiple high NA objectives without requiring separate large-scale optical paths
2Measurement precision
If multiple high numerical aperture microscope objectives are placed at significant angles to each other, then oblique plane microscopy can be achieved, but the sample preparation and mounting becomes complicated
Solution Approach 1:
The patent employs a single objective lens that serves multiple functions: it provides both illumination and detection capabilities, and can be used with various sample mounting methods including conventional microscope coverslips and multiwell plates. This universal objective design eliminates the need for specialized sample preparation procedures required by dual-objective systems
Solution Approach 2:
The patent merges the illumination and detection functions into a single objective lens system. By combining these functions and using a beam splitter to separate the illumination and detection beams, the system simplifies sample mounting requirements while maintaining the capability for oblique plane microscopy
3Speed
If remote objectives are used for scanning, then rapid remote scanning can be achieved, but the objectives must be placed at significant angles and their focal planes must intersect, which is difficult to achieve
Solution Approach 1:
The patent introduces a beam splitter as an intermediary element that mediates between the single objective lens and the detection camera. The beam splitter directs illumination beams through the objective to the sample and redirects emitted light from the sample to the camera, enabling rapid scanning without requiring complex angular arrangements of multiple remote objectives
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 'folded' OPM configuration achieves a more compact optical arrangement, enhances fluorescence collection efficiency, and allows for efficient and reliable 3D imaging through rapid scanning of the illumination sheet and field of view, without the need for complex axial adjustments of remote objectives.
Implementation Method 1
a first mirror located at said focal plane of the second objective lens, arranged to receive and reflect the beam of emitted light from the first objective lens
Implementation Method 2
the first mirror is also arranged to receive the beam of illumination from the illumination generator and to reflect the beam of illumination through the second objective lens
Implementation Method 3
a beam splitter disposed between the second objective lens and the second relay lens, the beam splitter being configured to: (i) direct the beam of illumination from the second objective lens to the second relay lens
Implementation Method 4
direct the beam of emitted light from the second relay lens to the second objective lens
Implementation Method 5
an illumination generator arranged to provide a beam of illumination through a first objective lens to illuminate or excite an oblique plane of a sample in use, wherein the first objective lens is also arranged to receive a beam of emitted light from the oblique plane of the sample in use
Data Source
AI summary
An optical arrangement for oblique plane microscopy, comprising: an illumination generator arranged to provide a beam of illumination through a first objective lens to illuminate or excite an oblique plane of a sample in use, wherein the first objective lens is also arranged to receive a beam of emitted light from the oblique plane of the sample in use; first and second relay lenses and a second objective lens sequentially arranged to receive the beam of emitted light from the first objective lens and to form, at the focal plane of the second objective lens, an intermediate image having a tilted plane conjugate to that of the oblique plane of the sample; a first mirror located at said focal plane of the second objective lens, arranged to receive and reflect the beam of emitted light; a third relay lens; and an image detector; wherein the second objective lens and the third relay lens are arranged to relay the intermediate image from the first mirror to the image detector; wherein the first minor is also arranged to receive the beam of illumination from the illumination generator and to reflect the beam of illumination through the second objective lens; and wherein the optical arrangement further comprises a beam splitter disposed between the second objective lens and the second relay lens, the beam splitter being configured to: (i) direct the beam of illumination from the second objective lens to the second relay lens, and thence to the first relay lens, the first objective lens and the sample; (ii) direct the beam of emitted light from the second relay lens to the second objective lens, and thence to the first mirror; and (iii) direct the reflected beam of emitted light from the second objective lens to the third relay lens and thence to the image detector. Also provided is a corresponding method of performing oblique plane microscopy.