Multiscale Spectral Nanoscopy Tracking
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current imaging methods are limited in their ability to observe molecular interactions and dynamics at the native length and time scales relevant to biological processes, particularly in three dimensions with high temporal resolution, due to resolution limits and slow acquisition times.
Innovation Solution
The Multiscale Spectral Nanoscopy (MSN) system combines a 2-photon or confocal laser scanning microscope with a particle-holding device and tracking module, enabling high spatial and temporal resolution tracking of particles across multiple scales, from sub-nm to multicellular levels, using a 3D piezo stage and spectroscopic capabilities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional confocal microscopy is used to achieve high spatial resolution, then imaging quality is improved, but acquisition speed deteriorates to ≤1 Hz which is too slow for real-time biological processes
Solution Approach 1:
The system segments the imaging task into two parallel pathways: a fast camera-based pathway for real-time tracking of selected particles at high temporal resolution, and a slower scanning microscopy pathway for comprehensive spectral and spatial mapping. This segmentation allows each pathway to be optimized for its specific function without compromising the other.
Solution Approach 2:
The invention adds a temporal dimension to the imaging system by implementing continuous real-time tracking alongside spatial spectral mapping. The fast camera captures temporal dynamics at high frame rates while the scanning microscope provides spatial context, creating a multi-dimensional observation space that simultaneously addresses both speed and resolution requirements.
2Measurement precision
If super-resolution methods like STED or PALM are used to achieve resolution below diffraction limit, then measurement precision is improved to 10-30 nm, but temporal resolution deteriorates requiring tens of seconds per image
Solution Approach 1:
The system segments the observation target into individually tracked particles that can be monitored continuously over time. By focusing the super-resolution capability on specific selected particles rather than attempting to image entire fields at super-resolution, the system achieves high temporal resolution for individual particles while maintaining spatial precision.
Solution Approach 2:
The system applies super-resolution tracking selectively to specific particles of interest rather than attempting to achieve super-resolution across the entire field of view. This partial application of super-resolution capabilities allows real-time tracking of individual particles at high temporal resolution while using standard resolution for contextual imaging.
3Productivity
If fast 3D imaging methods like SPIM are used to achieve high acquisition speed, then productivity is improved to 1 Hz or faster, but measurement precision deteriorates with axial resolution on the order of 5-500 nm
Solution Approach 1:
The system segments the imaging task by using the fast SPIM pathway for temporal tracking and the scanning microscopy pathway for axial resolution optimization. The scanning microscope's confocal or two-photon excitation provides superior axial sectioning and resolution for particles of interest, while SPIM handles the high-speed temporal component.
Solution Approach 2:
The system merges two complementary imaging modalities: fast 3D imaging (SPIM) for temporal resolution and scanning microscopy (confocal or two-photon) for axial resolution. By combining these approaches in a coordinated system where particles are first identified by fast imaging and then tracked with high axial resolution, the system achieves both high acquisition speed and measurement precision.
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
MSN provides the first system capable of covering extensive length and time scales, enabling real-time observation of molecular interactions and dynamics, including organelle trafficking and single-molecule behavior within the cellular context, with high precision and sensitivity.
Implementation Method 1
a 2-photon or confocal laser scanning microscope (LSM) configured to scan the sample
Implementation Method 2
detect fluorescent signals from the particle
Implementation Method 3
X-Y radiation-gathering components configured to detect deviations of the particle in an X-Y direction and Z radiation-gathering components configured to detect deviations of the particle in a Z direction
Implementation Method 4
a processor coupled to the X-Y and Z radiation gathering components, generate control signals configured to drive the stage X-Y and Z position controls to track the movement of the particle
Data Source
Figure 1~2
Figure 3A~3B
Figure 4~5
AI summary
A system and method for non-invasively tracking a particle in a sample is disclosed. The system includes a 2-photon or confocal laser scanning microscope (LSM) and a particle-holding device coupled to a stage with X-Y and Z position control. The system also includes a tracking module having a tracking excitation laser, X-Y and Z radiation-gathering components configured to detect deviations of the particle in an X-Y and Z directions. The system also includes a processor coupled to the X-Y and Z radiation gathering components, generate control signals configured to drive the stage X-Y and Z position controls to track the movement of the particle. The system may also include a synchronization module configured to generate LSM pixels stamped with stage position and a processing module configured to generate a 3D image showing the 3D trajectory of a particle using the LSM pixels stamped with stage position.