Implementation of Post-Processing Metrics for STORM Imaging
Objective
To integrate a suite of post-processing metrics into our STORM Image Processing GUI that will enable users to evaluate detailed fluorescence properties of localized molecules. This feature will enhance our tool's analytical capabilities, providing researchers with in-depth insights into their imaging data.
Requirements
Metrics to be Implemented:
On-Time/Total: Ratio of the time a molecule spends in the on state to the total observation period.
On-Time and Off-Time Average: Calculate the average duration of on and off states.
Standard Deviation in Localization: Measure the variability in the molecule’s position and quantify error from its possible true center.
Intensity Over Time: Track changes in fluorescence intensity throughout the observation period.
Duty Cycle: Calculate the ratio of on-time to off-time.
Instant Time of State Transition: Measure the duration from the highest intensity to the lowest intensity during a state transition.
Number of Photons Localized per Frame: Compute the average count of photons detected per frame.
Number of Overcounting Found per Frame: Identify incidences of detecting two particles closer together than what is defined by Abbe’s diffraction limit in a frame.
Switching Cycles: Determine the average number of on-off cycles a molecule undergoes before it permanently photobleaches.
Survival Fraction: Evaluate the proportion of fluorophores that remain capable of switching post-illumination compared to those permanently bleached.
Expandability: The system should be designed to easily incorporate additional metrics as new research and needs arise.
Goals
Develop Robust Algorithms: Create algorithms to compute each of the specified metrics from the processed image data.
Update GUI: Design and implement updates to the GUI to display these metrics in an intuitive and accessible manner.
Ensure Accuracy and Performance: Validate the accuracy of the metrics and optimize performance to handle large datasets efficiently.
Challenges
Data Complexity: Handling the complex data sets typical of STORM imaging and providing accurate, reliable metrics.
User Interface Design: Ensuring the GUI remains user-friendly and informative despite the addition of detailed and potentially complex metrics.
Milestones
Algorithm Design: Define and develop algorithms for each metric.
GUI Integration: Update the GUI to include new sections or panels for displaying metrics.
Feature Testing: Extensively test the feature with diverse datasets to ensure reliability and accuracy.
User Feedback: Collect and incorporate user feedback to refine the functionality.
Documentation: Update documentation to include detailed descriptions of each metric and instructions for use.
Expected Outcomes
A comprehensive set of post-processing metrics that will provide STORM researchers with deeper insights into their imaging experiments.
An enhanced user experience in our GUI, making it a more powerful tool for scientific analysis.
Implementation of Post-Processing Metrics for STORM Imaging
Objective
To integrate a suite of post-processing metrics into our STORM Image Processing GUI that will enable users to evaluate detailed fluorescence properties of localized molecules. This feature will enhance our tool's analytical capabilities, providing researchers with in-depth insights into their imaging data.
Requirements
Metrics to be Implemented:
Expandability: The system should be designed to easily incorporate additional metrics as new research and needs arise.
Goals
Challenges
Milestones
Expected Outcomes
Visuals and Documentation