Multi-Plane Imaging System Using Temporal Multiplexing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Two-photon laser scanning microscopy is limited by the raster scanning rate, restricting the number of regions of interest that can be imaged sequentially with sufficient temporal resolution, which hinders faster imaging and broader volume coverage in neuroscience applications.
Innovation Solution
A system employing multiple remote focusing units and temporal division-multiplexing techniques to enable simultaneous multi-plane imaging by generating and demultiplexing temporally interleaved excitation pulses, allowing independent axial positioning of imaging planes and shared lateral positioning, thereby increasing imaging throughput.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If raster scanning is used for imaging, then image quality and resolution are maintained, but imaging speed is limited
Solution Approach 1:
The patent segments the imaging process by dividing it into multiple independent focal planes that can be imaged simultaneously. Instead of sequentially scanning through Z-positions, the system creates multiple remote focusing units, each targeting a different focal plane, allowing parallel acquisition of images from multiple depths at once, thereby dramatically increasing imaging throughput while maintaining resolution
Solution Approach 2:
The patent transitions from two-dimensional lateral scanning to three-dimensional simultaneous multi-plane imaging by adding the axial dimension as an additional imaging plane. Multiple focal planes at different Z-positions are imaged concurrently, converting a sequential Z-stack acquisition into a parallel multi-plane process, thus improving both speed and productivity
2Measurement precision
If sequential imaging of multiple regions is performed, then comprehensive coverage is achieved, but temporal resolution is insufficient
Solution Approach 1:
The patent segments the imaging volume into multiple discrete focal planes that can be independently and simultaneously imaged. Each focal plane acts as an independent region of interest, allowing the system to capture temporal dynamics across multiple regions concurrently rather than sequentially, thereby improving both temporal resolution and the number of regions imaged
Solution Approach 2:
The patent enables continuous simultaneous imaging across multiple focal planes without interruption or sequential transitions. All focal planes are illuminated and detected concurrently, maintaining continuous useful action across the entire imaging volume, which preserves temporal resolution while increasing the number of regions captured
3Productivity
If multiple focal planes are imaged simultaneously, then imaging throughput increases, but signal separation becomes challenging
Solution Approach 1:
The patent employs periodic temporal modulation of the excitation signals directed at each focal plane, creating distinct temporal signatures for each plane. By alternating or modulating the excitation pulses in a periodic pattern, the system enables temporal demultiplexing of the emitted signals, making it easier to separate and identify signals from different focal planes while maintaining simultaneous imaging throughput
Solution Approach 2:
The patent applies polarization encoding to distinguish signals from different focal planes, analogous to using different colors. Each focal plane is excited with light of a specific polarization state, and the detection system is configured to selectively detect polarized emissions, enabling clear separation of signals from multiple planes without interference
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
The system effectively doubles the imaging throughput of two-photon microscopy, reducing cross-talk and maintaining signal quality, enabling simultaneous imaging from multiple focal planes with improved temporal resolution and larger field of view.
Implementation Method 1
a light source to generate an optical signal, the optical signal including a set of pulses at a first repetition rate
Implementation Method 2
an electro-optic modulator and a polarizing beam splitter to receive the optical signal and to generate a multiplexed optical signal
Implementation Method 3
a focusing unit to receive the multiplexed optical signal and to split the multiplexed optical signal into a set of n excitation signals... The objective and the focusing unit collectively focus each excitation signal of the set of n excitation signals on a different focal plane of the sample
Implementation Method 4
an objective to receive the set of n excitation signals and to illuminate the sample
Implementation Method 5
a detector to generate a response signal based on the emission from the sample and a switch to separate the response signal into a first emission signal corresponding to the first excitation signal and a second emission signal corresponding to the second excitation signal
Data Source
Figure 1
Figure 2
Figure 3~4
AI summary
A system includes a light source to generate an optical signal having a set of pulses at a first repetition rate. The system also includes a multiplexer circuit to generate a multiplexed optical signal from the optical signal n sets of pulses at a second repetition rate, where the n sets of pulses have different polarization states and are at the first repetition rate. The system also includes a focusing unit to split the multiplexed optical signal into n excitation signals to excite a sample. The system also includes an objective to receive the n excitation signals and to illuminate the sample. The objective and the focusing unit collectively focus each excitation signal of the n excitation signals on a different focal plane of the sample to generate a response signal. The system also includes a demultiplexer circuit to generate n emission signals based on the response signal.