3D Fluorescence Reconstruction in Non-Uniform Refractive Samples
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Solution Overview
Problem
Existing fluorescence imaging methods struggle with reconstructing the spatial distribution of fluorescence in non-uniform refractive index samples, particularly in biological tissues, often requiring invasive clarifying agents that disrupt sample integrity and cannot be used in real-time.
Innovation Solution
A non-invasive method for reconstructing three-dimensional fluorescence distribution using iterative algorithms that account for non-uniform refractive indices, employing beam propagation methods (BPM) to model light propagation and back-propagation through the object, with random phase assignments to voxels to handle incoherent fluorescence emissions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a clarifying agent is used to limit the influence of non-uniformity in refractive indices, then 3D images of fluorescence can be obtained without suffering from refractive index non-uniformity, but the method becomes invasive and does not allow real-time sample following
Solution Approach 1:
The patent replaces the mechanical/chemical approach of using clarifying agents with a computational/optical approach. Instead of physically altering the sample to eliminate refractive index variations, the system uses iterative algorithms that model light propagation through the actual non-uniform refractive index distribution, substituting physical correction with mathematical modeling to achieve accurate 3D fluorescence reconstruction without sample modification
Solution Approach 2:
The patent introduces a computational intermediary - an iterative algorithm that acts as a mediator between the measured 2D fluorescence images and the desired 3D fluorescence distribution. This algorithm processes the images by repeatedly modeling light propagation through the object's refractive index structure, effectively translating incomplete 2D measurements into accurate 3D reconstructions without requiring physical intervention in the sample
2Measurement precision
If iterative algorithms with beam propagation methods are used to model light propagation through non-uniform refractive indices, then accurate 3D fluorescence distribution can be reconstructed without clarifying agents, but the computational complexity and processing time increase
Solution Approach 1:
The patent implements a feedback mechanism through iterative algorithms that repeatedly model light propagation and compare predicted fluorescence distributions with actual measurements. The algorithm adjusts the 3D fluorescence distribution estimate based on the discrepancy between modeled and measured images, progressively converging toward an accurate reconstruction. This feedback loop enables the system to handle complex refractive index variations systematically
Solution Approach 2:
The patent performs preliminary actions by pre-characterizing the object's refractive index distribution through optical diffraction tomography or other measurement techniques before the main fluorescence reconstruction process. This preliminary mapping of the refractive index structure allows the iterative algorithm to use accurate propagation models from the outset, reducing the number of iterations needed and lowering overall computational complexity while maintaining high reconstruction accuracy
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 accurate reconstruction of fluorescence distribution without clarifying agents, maintaining sample integrity and allowing real-time analysis, despite non-uniform refractive indices.
Implementation Method 1
the object emitting fluorescence light under the effect of illumination in an excitation spectral band
Implementation Method 2
implementing an algorithm modelling propagation of light through the object, so as to estimate a complex amplitude of the light wave detected by each image voxel
Data Source
AI summary
The invention describes an iterative reconstructing method allowing a spatial distribution of fluorescence in an object to be obtained. The method comprises acquiring images of fluorescence in various planes at various depths in the object, so as to form a three-dimensional acquired image. It comprises an iterative reconstructing algorithm with, in each iteration, an initial fluorescence distribution or a fluorescence distribution resulting from a preceding iteration being taken into account, and the fluorescence light wave propagating through the object being simulated, so as to obtain a reconstruction of the acquired image. The acquired image, or a differential image corresponding to a comparison between the acquired image and the reconstructed image, is then back-propagated through the object, so as to update the fluorescence distribution. FIG. 5B.