Chromatic Optical System for Fluorescence and Defocused Imaging
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Observing transparent or translucent biological samples using fluorescence microscopy is challenging as the visible image is not usable for precise structural information, leading to difficulties in obtaining accurate sample structure details.
Innovation Solution
A method and device utilizing a bimodal imaging approach with a chromatic optical system, where a sample is illuminated with two distinct spectral bands to capture a focused fluorescence image and a defocused image, allowing for holographic reconstruction to obtain structural information without moving the sample or optical system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
If fluorescence microscopy is used to observe transparent or translucent samples, then fluorescence information can be obtained, but the visible image is not usable for obtaining precise structural information
Solution Approach 1:
The patent segments the imaging process into two distinct spectral channels: fluorescence imaging for molecular information and visible light defocused imaging for structural information. This segmentation allows each channel to optimize for its specific purpose while avoiding the limitation where fluorescence alone cannot provide adequate structural details of transparent samples.
Solution Approach 2:
The patent introduces a defocused imaging dimension by intentionally placing the sample plane outside the focal plane of the optical system. This creates a defocused visible image that encodes phase and structural information of transparent samples, adding a new dimension of information that complements the fluorescence data.
2Loss of information
If the sample is defocused to obtain structural information, then phase shift information can be obtained, but the fluorescence image quality deteriorates
Solution Approach 1:
The patent segments the imaging function by using a chromatic optical system where different wavelengths focus at different positions. The fluorescence spectral band is focused on the sample plane for sharp fluorescence imaging, while the visible spectral band is intentionally defocused to capture phase information, allowing both functions to operate optimally simultaneously.
Solution Approach 2:
The patent changes the focal plane parameter differently for different spectral bands. By exploiting chromatic aberration, the system sets the focal plane for fluorescence wavelengths at the sample position while setting the focal plane for visible wavelengths at a different position, thereby achieving both focused fluorescence and defocused visible imaging.
3Adaptability or versatility
If the sample or optical system is moved to switch between imaging modalities, then both fluorescence and structural images can be obtained, but the device complexity increases
Solution Approach 1:
The patent makes the chromatic optical system universal by designing it to perform both fluorescence imaging and defocused visible imaging simultaneously without requiring mechanical movement. The single optical system handles multiple imaging modalities by exploiting wavelength-dependent focal plane differences, eliminating the need for separate optical paths or moving components.
Solution Approach 2:
The patent replaces mechanical moving systems with an optical solution. Instead of physically moving the sample or optical components to switch between focused and defocused modes, the system uses chromatic aberration to achieve different focal states for different wavelengths, substituting mechanical actuation with optical physics.
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 the acquisition of both fluorescence and structural images of transparent or translucent samples, providing precise structural information without the need for adjusting focal planes or moving components, thus overcoming the limitations of traditional microscopy methods.
Implementation Method 1
the optical system being such that: in the fluorescence spectral band, the object focal plane of the optical system coincides with the plane sample; in the second spectral band, the object focal plane of the optical system is shifted with respect to the sample plane
Implementation Method 2
the sample comprising a fluorescent agent capable emitting a fluorescence light wave, in a fluorescence spectral band, when illuminated by an excitation light wave, in an excitation spectral band
Data Source
Figure 1A~1B
Figure 1C~1D
Figure 2A~3D
AI summary
The invention is a method for observing a fluorescent sample (10), extending along a plane (P10), called the sample plane, the sample comprising a fluorescent agent (10f) capable of emitting a fluorescence light wave (Δf), in a fluorescence spectral band (Δλf), when illuminated by an excitation light wave (Δe), in an excitation spectral band (Δλe), the method comprising the following steps: a) illumination of the sample (10) using a first light source (11), according to a first illumination spectral band (Δλ1), according to the excitation spectral band (Δλe) and acquisition of a first image of the sample (I1), in the fluorescence spectral band, using an image sensor (30);b) illumination of the sample (10) using a second light source (12), according to a second spectral band (Δλ2), outside the fluorescence spectral band, and acquisition of a second image of the sample (I2), in the second spectral band, using the image sensor; the image sensor (30) being coupled to an optical system (20) such that: - in the fluorescence spectral band (Δλf), the object focal plane of the optical system (P1) coincides with the plane of the sample (P10); - in the second spectral band, the object focal plane of the optical system (P2) is offset with respect to the plane of the sample (P10), the offset being greater than 20 µm.