Dynamic Light-Sheet Light-Field Microscopy for Higher Image Contrast
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Solution Overview
Problem
Light-field microscopy struggles to discriminate emission light generated away from the focus, leading to reduced image contrast and collection efficiency, particularly in thick biological samples, and existing methods to improve contrast often compromise the achievable volume rate.
Innovation Solution
A dynamic light sheet is generated by modulating the intensity distribution of illumination radiation, altering its angle of incidence multiple times, and using a microlens array to capture partial image representations, which are then evaluated for location and angle information to reduce unwanted background signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If a continuous light sheet scan is used to illuminate the sample, then the illumination covers the entire sample field, but the excitation light leaves the sample field after a certain propagation length and backscattering is not implemented over the entire numerical aperture, resulting in reduced collection efficiency and poor image contrast
Solution Approach 1:
The patent applies local quality by using a light sheet that selectively illuminates only the focal plane of the sample at a time, rather than the entire sample field. This localized illumination approach ensures that excitation light remains confined within the detection numerical aperture, maximizing backscattering collection efficiency and improving image contrast while maintaining adequate illumination intensity in the illuminated region
Solution Approach 2:
The patent employs periodic action by scanning the light sheet through different positions and angles systematically. The light sheet is moved in a controlled periodic manner across the sample, with each position captured by the camera, allowing comprehensive sample illumination over time while maintaining optimal local illumination conditions at each moment
2Measurement precision
If a triggered light-sheet scan is used to capture images at each light sheet position, then unwanted background signals can be removed by calculation, but the achievable volume rate is significantly reduced
Solution Approach 1:
The patent uses periodic action by implementing a triggered light-sheet scan where the camera is synchronized to capture images at each light sheet position. The light sheet scans periodically through the sample, and the camera triggers at each position to capture the illuminated region. This periodic scanning and capturing approach enables background signal removal through calculation while maintaining a manageable volume rate by optimizing the scanning speed and camera trigger timing
Solution Approach 2:
The patent applies continuity of useful action by ensuring that the light sheet scanning and camera capture operate continuously in a synchronized manner. Rather than stopping at each position, the system maintains continuous scanning and capturing, allowing efficient data acquisition that preserves volume rate while still enabling background removal through the periodic triggering scheme
3Measurement precision
If structured illumination is used to eliminate out-of-focus background light, then image contrast is improved, but the loss of volume rate increases due to requiring multiple image recordings
Solution Approach 1:
The patent applies local quality by implementing structured illumination that modulates light intensity at different positions within the light sheet. By creating structured patterns (such as lines or regions of varying intensity) in the illuminated area, the system can selectively enhance contrast for in-focus structures while minimizing the impact on volume rate, as the structuring is applied locally rather than requiring multiple full-sample scans
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
This approach enhances image contrast while maintaining a high volume rate by selectively illuminating sample sections, reducing background noise through structured illumination and computational methods, suitable for both frequency and spatial domain light-field imaging.
Implementation Method 1
provision of an illumination radiation from at least one light source
Implementation Method 2
A detection radiation coming from the sample space is collected by means of the common objective
Implementation Method 3
A plurality of partial image representations are generated in a detection plane by means of a microlens array arranged in the detection beam path
Implementation Method 4
fluorescence markers or fluorophores serve marking purposes in this context. As a result of illumination with radiation at an excitation wavelength, a fluorescence radiation, as detection radiation, can be excited
Data Source
AI summary
The invention relates to a light-field microscope and a light-field microscopy image capturing method, including the steps of providing an illumination radiation from at least one light source; modulating the illumination radiation by virtue of adjusting the intensity distribution thereof; generating a dynamic light sheet in a sample space, in which a sample to be imaged can be arranged; scanning a region of the sample to be imaged using the light sheet; collecting a detection radiation and detecting image signals using light-field technology. The illumination radiation is modulated by virtue of the light source being controlled and an intensity of the illumination radiation provided by the light source being altered, especially in a controlled fashion over time. Image recordings of the sample are triggered for different positions of the light sheet.