Random Intensity Illumination Microscopy for Optical Sectioning
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Solution Overview
Problem
Current optical microscopy techniques face limitations in providing rapid, full-field image acquisition with high-quality sectioning at depth while being cost-effective and safe for in-vivo measurements, particularly due to the need for expensive equipment, fluorescence staining, and artifacts like streaking in images.
Innovation Solution
An optical microscopy system utilizing an incoherent light source and a static speckle diffusion pattern, rotatable about an axis, to image samples using reflected optical signals, which includes an incoherent light source, diffusers for creating spatially random diffusion patterns, a beam splitter, an objective lens, and an image capture device with digital image processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If confocal microscopy is used to section individual planes and eliminate out of focus light, then image sectioning quality is improved, but equipment cost and complexity increase due to required point scanning equipment
Solution Approach 1:
The patent replaces the mechanical point-scanning system with a static random diffusion pattern illuminated by an incoherent light source. Instead of mechanically scanning points through a pinhole, the system projects a random pattern that provides optical sectioning through incoherent illumination, eliminating the need for complex scanning mechanisms while maintaining sectioning capability
Solution Approach 2:
The patent uses inexpensive incoherent light sources and simple static diffusion patterns instead of expensive confocal scanning equipment. The random diffusion pattern can be generated by simple optical elements rather than complex mechanical scanners, significantly reducing system cost and complexity
2Measurement precision
If dynamic speckle illumination is used to provide depth information, then axial resolution is improved, but the number of images required increases with depth reducing sectioning quality
Solution Approach 1:
The patent pre-generates a static random diffusion pattern that is projected onto the sample before imaging. This preliminary pattern projection enables depth information extraction from a single image or minimal images, eliminating the need to capture multiple images as depth increases in dynamic speckle illumination
Solution Approach 2:
The patent uses a rotatable diffuser that can be dynamically rotated to change the random diffusion pattern between images. This dynamic rotation provides temporal variation in the illumination pattern, enabling depth information extraction while maintaining rapid acquisition
3Measurement precision
If dynamic speckle illumination is used with fluorescence, then depth information is obtained, but photobleaching and potential harm to in-vivo specimens occur
Solution Approach 1:
The patent changes the illumination parameter from coherent laser light to incoherent light from an extended source. This parameter change eliminates speckle patterns while maintaining the random diffusion effect, allowing depth information extraction without requiring fluorescence excitation, thereby avoiding photobleaching and specimen damage
Solution Approach 2:
The patent extracts the depth information extraction capability from the fluorescence requirement. By using incoherent illumination with random diffusion patterns, the system obtains axial sectioning and depth information through reflectance imaging alone, removing the need for fluorescent staining and its associated harms
4Measurement precision
If structured illumination with constant spatial frequency is used, then image sectioning is achieved, but the system requires expensive and elaborate equipment
Solution Approach 1:
The patent changes the illumination pattern parameter from a constant spatial frequency structured pattern to a random diffusion pattern with multiple spatial frequencies. This parameter change simplifies the optical system by eliminating the need for precise pattern generation and phase shifting mechanisms, reducing equipment complexity while maintaining sectioning capability
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 rapid, high-quality image acquisition with reduced costs and safety for in-vivo measurements, minimizing artifacts and extending the range of applications for optical microscopy, including biomedical and healthcare uses by improving diagnostic times and patient care.
Implementation Method 1
an incoherent light source configured to provide an incoherent light signal
Implementation Method 2
at least two diffusers coupled to the incoherent light signal and configured to provide a diffused light signal having spatially random diffusion patterns
Implementation Method 3
an objective lens coupled to the first light signal and configured to focus the first signal on a sample object to be imaged to provide a reflected signal
Implementation Method 4
a beam splitter coupled to the diffused light signal and configured to provide a first light signal and a second light signal
Implementation Method 5
an image capture device, coupled to the beam splitter so as to receive the first light signal and the reflected light signal
Data Source
AI summary
An optical imaging system and method for performing random intensity illumination microscopy is disclosed. The system includes an incoherent signal light source, at least two diffusers having spatially random diffusion patterns, an image capture device that receives a reflected light signal from an object to be imaged, and a processor configured to perform digital image processing of the reflected signal. The method comprises acts of providing an incoherent light signal, diffusing the incoherent light signal with at least two diffusers having spatially random diffusion patterns to provide a diffused light signal, splitting the diffused light signal to provide a first light signal and a second light signal, reflecting the first light signal from a specimen to provide a reflected light signal, collecting the reflected light signal and the second light signal with an image capture device and processing the collected images to determine reflectance.