Confocal Light Scattering Spectroscopy for Sub-Rayleigh Detection
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Solution Overview
Problem
Current methods for detecting microscopic objects, such as confocal microscopy, two-photon fluorescence, light scattering spectroscopy, and near-field optical microscopy, have limitations including diffraction limits, requirement for fluorophores, complexity with densely packed objects, and limited depth of field, which restrict their effectiveness in various applications.
Innovation Solution
A confocal light scattering spectroscopy (CLSS) system combining broadband illumination with a confocal optical system and spectral separation, allowing for the detection of objects with dimensions below the Rayleigh limit and providing high accuracy in three dimensions, using a two-dimensional detector and spectral separation device to process scattered light over a large angle.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If confocal microscopy is used to detect microscopic objects, then spatial resolution is improved, but detection capability is limited by diffraction to objects greater than the Rayleigh limit
Solution Approach 1:
The patent combines confocal microscopy with light scattering spectroscopy to create a hybrid system that merges the spatial resolution advantages of confocal microscopy with the diffraction-limit-free detection capabilities of light scattering spectroscopy, enabling detection of objects below the Rayleigh limit while maintaining spatial resolution
Solution Approach 2:
The system performs multiple functions simultaneously: it provides confocal imaging for spatial localization and light scattering spectroscopy for size measurement, making the system versatile for both imaging and characterization of microscopic objects below the diffraction limit
2Measurement precision
If two-photon fluorescence is used to detect microscopic objects, then detection sensitivity is improved, but the method requires fluorophores that may cause artifacts or destroy the object
Solution Approach 1:
The system uses the object's own light scattering properties as the measurement mechanism, requiring no external fluorophores or labels. The object serves itself by scattering light, eliminating the need for potentially harmful fluorescent tags while maintaining detection sensitivity
Solution Approach 2:
The patent extracts the detection mechanism from fluorophore-dependent fluorescence and replaces it with fluorophore-independent light scattering spectroscopy, removing the requirement for potentially damaging fluorophores while preserving detection capabilities
3Adaptability or versatility
If light scattering spectroscopy is used to detect densely packed objects, then detection capability is maintained, but the method becomes complicated due to multiple light scattering
Solution Approach 1:
The confocal pinhole extracts only the light scattered from the specific focal volume, removing out-of-focus scattered light that would complicate the measurement. This isolation of the confocal volume simplifies the analysis of densely packed objects by eliminating multiple scattering paths from other regions
4Measurement precision
If near-field optical microscopy is used to achieve high resolution, then spatial resolution below the Rayleigh limit is improved, but depth of field becomes very limited
Solution Approach 1:
The confocal system maintains continuous useful action by collecting light scattered over a large angular range (greater than 10 degrees) from the confocal volume, providing robust signal collection that maintains detection capability across the available depth of field without the severe depth limitations of near-field microscopy
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 non-invasive detection of microscopic objects with dimensions as small as 0.5 micrometers, achieving high accuracy in size measurement to within 5-10 nanometers and providing capabilities not available in conventional methods, such as detecting objects below the Rayleigh limit and characterizing objects in three dimensions.
Implementation Method 1
light scattered from a confocal volume on the object is received
Implementation Method 2
a spectral separation device configured and arranged to receive the scattered light from the object and to direct at least a portion of the scattered light onto the two-dimensional detector, the spectral separation device spectrally separating the portion of the light
Data Source
AI summary
Apparatus including a broadband illumination source and a confocal optical system. The confocal optical system is configured and arranged to receive a portion of light projected onto an object by the broadband illumination source. The apparatus can include an illumination source, a confocal optical system, and at least one detector configured and arranged to receive angularly separated light corresponding to a confocal volume. There is also provided a light scattering spectroscopic device including a broadband illumination source, a two-dimensional detector, and a spectral separation device configured and arranged to receive scattered light from an object and to direct at least a portion of the scattered light onto the two-dimensional detector. The method and apparatus can combine confocal microscopy techniques with light scattering spectroscopy techniques to create a confocal light scattering spectroscopy (CLSS) system.