Nucleic Acid Complexes for Simultaneous Lysosomal pH and Ca2+ Sensing
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Solution Overview
Problem
Current methods are unable to chemically resolve lysosome populations in live cells due to the pH sensitivity of Ca2+ indicators, which impedes the quantification of lumenal Ca2+ and Cl− concentrations, crucial for understanding lysosome function and disease pathology.
Innovation Solution
Development of nucleic acid complexes, such as CalipHluorLy and ChloropHore, that include Ca2+ and Cl− fluorophores crosslinked with complementary nucleic acid molecules, allowing simultaneous determination of pH, Ca2+ concentration, or Cl− concentration by measuring signal intensity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If pH-sensitive Ca2+ indicators are used to measure lumenal Ca2+ concentration, then Ca2+ quantification becomes possible, but measurement accuracy deteriorates due to pH sensitivity interfering with the readings
Solution Approach 1:
The invention divides the measurement function into two separate probes: one probe specifically for pH measurement and another for Ca2+ concentration measurement. This segmentation allows each probe to be optimized for its specific target without interference from the other parameter, resolving the contradiction between Ca2+ measurement accuracy and pH sensitivity interference.
Solution Approach 2:
The invention introduces pH correction as an intermediary step in the measurement process. By first measuring pH and then using this information to correct the Ca2+ indicator readings, the system eliminates the harmful effect of pH sensitivity on Ca2+ measurement accuracy.
2Loss of information
If multiple lysosome populations are chemically resolved, then understanding of lysosome function improves, but device complexity increases due to need for multiple measurement parameters
Solution Approach 1:
The invention creates a universal measurement platform that can detect multiple lysosome parameters (pH, Ca2+, Cl−) using the same basic probe structure and imaging methodology. This multi-functionality allows comprehensive chemical resolution of lysosome populations without requiring separate complex systems for each parameter.
Solution Approach 2:
The invention combines multiple measurement capabilities into integrated probe molecules that can simultaneously report on pH, Ca2+, and/or Cl− concentrations. By merging these functions into single probes, the system reduces overall complexity while maintaining the ability to resolve multiple lysosome populations based on their chemical compositions.
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
Enables accurate and efficient mapping of lysosomal pH and Ca2+ or Cl− concentrations with single-endosome resolution, facilitating the study of lysosome function and disease progression.
Implementation Method 1
Ca2+ and Cl− fluorophores crosslinked with complementary nucleic acid molecules, allowing simultaneous determination of pH, Ca2+ concentration, or Cl− concentration by measuring signal intensity
Data Source
AI summary
This disclosure relates to methods for determining pH and also calcium (Ca2+) concentration or chloride (Cl−) concentration in biological samples. More particularly, this disclosure relates to methods capable of simultaneously determining pH and Ca2+ concentration, or pH and Cl− concentration using nucleic acid complexes.