Liquid Fluorescent Dye Concentrate for Virus-Size Particle Flow Cytometry
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Solution Overview
Problem
Flow cytometry for analyzing virus-size particles faces challenges in precision and repeatability due to self-quenching effects from fluorogenic dye aggregates and limited stability of stain formulations, leading to inconsistent results and high blank particle counts.
Innovation Solution
A liquid dye concentrate with multiple fluorogenic dyes in a liquid medium, including a disaccharide and a polar organic solvent like DMSO, is used to reduce dye aggregation and improve stability, providing a stable and effective staining solution for flow cytometry evaluation of virus-size particles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorogenic dyes are used for staining virus-size particles, then fluorescent signal detection is enabled, but self-quenching effects from dye aggregates reduce measurement precision
Solution Approach 1:
The patent introduces a disaccharide (such as trehalose, sucrose, or lactose) as an intermediary substance in the liquid staining formulation. This disaccharide acts as a mediator that prevents fluorogenic dye molecules from aggregating into self-quenching clusters, thereby maintaining fluorescent signal intensity and measurement precision without requiring dry powder mixtures or complex reconstitution procedures.
2Ease of operation
If multiple fluorogenic dyes are mixed in aqueous solution, then simultaneous staining is achieved, but formulation stability deteriorates over time
Solution Approach 1:
The patent changes the chemical parameters of the staining formulation by incorporating a disaccharide into the aqueous liquid medium. This parameter change (adding disaccharide at specific concentrations) fundamentally alters the physical-chemical environment to prevent dye aggregation and maintain formulation stability over extended periods, enabling ready-to-use liquid formulations with shelf lives comparable to dry powder mixtures.
3Duration of action of stationary object
If dry powder dye mixtures are used, then shelf life is extended, but reconstitution complexity increases
Solution Approach 1:
The patent extracts the disaccharide stabilizing component from complex dry powder mixture systems and incorporates it directly into ready-to-use liquid formulations. This eliminates the need for separate reconstitution steps with organic solvents like acetonitrile or DMSO, while maintaining extended shelf life and preventing dye aggregation in the final staining solution.
4Ease of operation
If fluorogenic dyes are stored in liquid form, then ease of use is improved, but aggregation and blank particle counts increase
Solution Approach 1:
The disaccharide in the liquid formulation serves as a protective intermediary that surrounds and separates fluorogenic dye molecules, preventing them from aggregating into self-quenching clusters. This maintains the dyes in a stable, dispersed state in liquid form, eliminating blank particle counts while preserving the ease of use associated with liquid formulations.
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 solution significantly reduces blank particle counts and enhances the precision and repeatability of flow cytometry results by minimizing dye aggregation and maintaining fluorogenic dye stability, offering a shelf-life comparable to dry powder mixtures without the complexity of reconstitution.
Implementation Method 1
a liquid dye concentrate comprising a plurality of different fluorogenic dyes in a liquid medium
Implementation Method 2
self-quenching effects from fluorogenic dye aggregates
Implementation Method 3
fluorescent emission signature for detection when bound to a virus-size particle
Data Source
AI summary
A kit and method for flow cytometry include a liquid dye concentrate for fluorescent staining of virus-size particles with a plurality of fluorogenic dyes in a liquid medium. The liquid dye concentrate includes a plurality of fluorogenic dyes and one or both of (i) the liquid medium comprising a liquid mixture including water and liquid phase organic material and (ii) disaccharide dissolved in the liquid medium.