Adaptive Depth-of-Field Control for Light-Sheet Photon Collection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Light-sheet microscopy is limited by the tradeoff between high numerical aperture objectives' short depth of field and the thicker excitation light-sheets used for large field-of-view imaging, leading to lost fluorescence signal photons, reduced signal-to-noise ratio, and compromised imaging speed and contrast.
Innovation Solution
Adjusting the depth of field (DOF) of the detection objective to match or exceed the thickness of the excitation light-sheet, using adaptive optical elements to capture a greater amount of fluorescence signal photons.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a high numerical aperture (NA) objective lens is employed for detection, then spatial resolution and light collection are improved, but depth of field becomes short causing loss of fluorescence signal photons
Solution Approach 1:
The patent employs adaptive optical elements (deformable mirror, spatial light modulator) that dynamically adjust the depth of field of the detection objective to match the thickness of the excitation light-sheet. This dynamic adaptation allows the system to optimize photon collection by extending the detection depth of field when imaging thicker light-sheets, thereby capturing fluorescence photons that would otherwise be lost due to the inherently short depth of field of high-NA objectives.
2Measurement precision
If a high numerical aperture (NA) objective lens is employed for detection, then signal-to-noise ratio is improved, but imaging speed is compromised due to short depth of field
Solution Approach 1:
The adaptive optical elements enable real-time adjustment of the detection depth of field to match the light-sheet thickness, allowing the system to maintain high signal-to-noise ratio while capturing photons from the entire thickness of the light-sheet. This dynamic optimization improves imaging speed by eliminating the need for multiple focal plane acquisitions that would be required with a fixed short depth of field.
3Quantity of substance
If the depth of field of the detection objective is extended to capture more photons, then signal-to-noise ratio and imaging speed are improved, but axial resolution deteriorates
Solution Approach 1:
The system dynamically adjusts the depth of field to match the light-sheet thickness rather than using a fixed extended depth of field. This dynamic matching ensures that the detection depth of field is extended only to the extent necessary to capture photons from the illuminated volume, thereby improving photon collection while minimizing the degradation of axial resolution that would occur with excessive depth of field extension.
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
Increases signal-to-noise ratio, improves image contrast, and enhances imaging speed by capturing more fluorescence photons while reducing photo damage and bleaching.
Implementation Method 1
illuminating a sample with an excitation light-sheet; receiving, at an imaging sensor, fluorescence signal data via the detection objective
Implementation Method 2
at least one adaptive optical element may be positioned at, or close to, any conjugate plane of the detection objective or one or more optics in the detection system. During imaging, the at least one adaptive optical element may adjust a DOF of the detection objective
Data Source
AI summary
Systems and methods are provided for increasing photon collection during imaging with a light-sheet fluorescence microscope. In one example, one or more adaptive optical elements may be positioned in a detection light path between a detection objective and an imaging sensor. A depth of field of the detection objective is adjusted as a function of a thickness of an excitation light-sheet used to illuminate a sample during imaging. As a result, the detection objective captures more fluorescence photons generated by light-sheet excitation.