Super-resolution Imaging Agent with Diffraction-limited Preview
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Solution Overview
Problem
Current super-resolution immunofluorescence techniques, such as PAINT-based Super-resolution Immunofluorescence (PSRIF), lack a preview mechanism, leading to inefficiencies in resource utilization and time as users cannot assess sample quality before initiating the lengthy imaging process, potentially resulting in wasted time and resources due to inadequate or incorrect samples.
Innovation Solution
The development of imaging agents comprising a target recognition moiety with a non-transiently bound observable moiety and a docking moiety capable of transient binding, allowing for a diffraction-limited preview of the sample before super-resolution imaging, enabling assessment and optimization of sample quality and composition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PSRIF collects multiple images over a period of time to achieve super-resolution imaging, then the resolution is improved beyond the diffraction limit, but the time required and resources consumed increase significantly
Solution Approach 1:
The patent applies preliminary action by performing a diffraction-limited preview imaging step before the super-resolution imaging process. This preview captures the sample at lower resolution but provides essential information about sample quality, cell type presence, and staining effectiveness, allowing users to assess whether the sample is suitable for the time-consuming super-resolution imaging process.
Solution Approach 2:
The imaging process is segmented into two distinct phases: a preliminary diffraction-limited preview phase and a subsequent super-resolution imaging phase. This segmentation allows the total process to be divided such that the expensive, time-consuming super-resolution imaging is only performed when necessary, based on the outcome of the quick preview assessment.
2Measurement precision
If PSRIF collects multiple images over an extended period to reconstruct the final image, then super-resolution is achieved, but experimental resources and computing time are wasted if the sample is inadequate or incorrect
Solution Approach 1:
The patent performs preliminary assessment actions through diffraction-limited preview imaging that evaluates sample adequacy, including checking for desired cell types in the field of view and assessing staining quality. This preliminary evaluation prevents waste of experimental resources and computing time by identifying unsuitable samples before committing to the resource-intensive super-resolution imaging process.
3Reliability
If PSRIF performs comprehensive sample assessment before imaging, then sample quality can be evaluated, but the overall process time increases
Solution Approach 1:
The assessment process is segmented into a rapid diffraction-limited preview phase that provides essential quality information quickly, followed by the optional super-resolution imaging phase. This segmentation enables reliable sample quality assessment without significantly increasing total process time, as the preview can be performed rapidly and determines whether further imaging is warranted.
Solution Approach 2:
The imaging workflow incorporates periodic assessment through the preview step that is performed before the main super-resolution imaging. This periodic check allows for quality evaluation at a strategic point in the process, enabling early termination if the sample is inadequate and preventing unnecessary extension of the total process time.
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 a preview phase that allows for the evaluation of sample quality and composition, optimizing resource allocation and reducing the risk of wasted time and resources by ensuring the sample is suitable for super-resolution imaging, thereby improving the efficiency of the imaging process.
Implementation Method 1
the observable moiety non-transiently bound to the target recognition moiety is a signal-emitting moiety
Implementation Method 2
the docking moiety is capable of transiently binding at least one observable moiety
Data Source
AI summary
An imaging agent comprises (a) at least one target recognition moiety; (b) at least one observable moiety non-transiently bound to the target recognition moiety, and (c) at least one docking moiety bound to the target recognition moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety. In some embodiments, the at least one target recognition moiety is an antibody or antigen binding fragment thereof. A method of performing super-resolution imaging on a sample comprises (a) contacting the sample with at least one imaging agent comprising (i) a target-recognizing molecule non-transiently attached to at least one observable moiety and (ii) a target-recognizing molecule attached to at least one docking moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety; (b) imaging the target-recognizing molecule non-transiently bound to at least one observable moiety; and (c) providing at least one observable moiety capable of transiently binding to the docking moiety and imaging the observable moiety transiently bound to the docking moiety.