PALM Microscopy Dynamic Exposure Control
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Solution Overview
Problem
Current PALM microscopy methods are time-consuming and require extensive data processing to achieve high spatial resolution, with image acquisition times often taking several minutes to several hours due to the need for multiple individual images and complex computational procedures.
Innovation Solution
The method involves evaluating individual images to optimize parameters dynamically during the acquisition process, allowing for controlled adjustments to variables such as filtering, excitation power, and detection settings to enhance image recording speed and quality, thereby reducing the overall time required for image acquisition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple individual images are recorded and combined to achieve high spatial resolution beyond the diffraction limit, then measurement precision is improved, but loss of time increases due to extensive data processing and long acquisition times
Solution Approach 1:
The patent segments the image acquisition process into multiple individual images, each capturing a subset of activated marker molecules. By recording multiple such images and combining them computationally, the system achieves high spatial resolution beyond the diffraction limit. Each individual image contains less data, but the collection of images provides comprehensive information for super-resolution reconstruction.
Solution Approach 2:
The patent employs preliminary action by activating only a subset of marker molecules in each imaging cycle rather than all molecules simultaneously. This selective activation is achieved through controlled switching signals that prime specific molecules for fluorescence emission, ensuring they are activated before the detection cycle begins. This preliminary selection reduces the number of molecules contributing to each individual image, thereby reducing total acquisition time while maintaining resolution through the accumulation of multiple such images.
2Measurement precision
If the number of individual images recorded is increased to improve localization accuracy, then measurement precision is improved, but productivity decreases due to longer acquisition times
Solution Approach 1:
The patent implements dynamics by adaptively adjusting the switching signal intensity and the number of individual images recorded based on real-time evaluation of image quality and marker molecule distribution. The system dynamically optimizes parameters such as excitation power, detection sensitivity, and acquisition cycles to achieve sufficient localization accuracy with minimal imaging steps, thereby improving productivity while maintaining precision.
Solution Approach 2:
The patent employs feedback mechanisms where individual recorded images are evaluated to determine whether additional images are needed for sufficient localization accuracy. The system uses this feedback to adjust the number of acquisition cycles, switching signal intensity, and other parameters, stopping the acquisition process once the required precision is achieved. This feedback-driven approach prevents unnecessary imaging steps and optimizes the balance between accuracy and speed.
3Measurement precision
If complex computational procedures are used to process multiple individual images, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent uses copying by creating multiple copies of the sample state at different time points, where each copy corresponds to an individual image containing a subset of activated marker molecules. Instead of processing a single complex image, the system works with multiple simpler copies that can be independently processed and then combined. This copying approach simplifies the computational task for each individual image while achieving high resolution through the synthesis of multiple copies.
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
This approach enables faster image acquisition and reduces the time needed to generate high-resolution images, allowing for more efficient use of sensitive detectors and cameras while maintaining or improving the accuracy of marker molecule localization beyond the diffraction limit.
Implementation Method 1
marker molecules that can be activated with a switching signal in such a way that they can only be excited to emit specific luminescence radiation in the activated state
Implementation Method 2
luminescence is understood here as a generic term for phosphorescence and fluorescence, ie it covers both processes
Implementation Method 3
luminescence is understood here as a generic term for phosphorescence and fluorescence, ie it covers both processes
Data Source
Figure 1~2
Figure 3~4(d)
Figure 5
AI summary
The invention relates to a method for PAL microscopy, wherein individual images are combined into a total image and there is a regulation of the exposure of the individual images in that at least a part or a group of the individual images are evaluated and at least one variable of the individual image exposure is modified for subsequent individual image exposures.