Fluorophore Wavelength Optimization via Signal-to-Background Analysis

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Conventional spectrofluorometry methods require manual setting and scanning of excitation-emission wavelength pairs, which are time-consuming and prone to error, often resulting in suboptimal sensitivity due to improper selection of bandpasses, leading to crosstalk or non-optimal intensity settings.

Innovation Solution

A computer-implemented system that automatically determines the optimal excitation-emission wavelength pair for a fluorophore by scanning both the fluorophore and a blank sample across specified wavelength ranges, calculating signal-to-background ratios, and identifying the maximum ratio to achieve optimal sensitivity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If manual setting and scanning of excitation-emission wavelength pairs is used, then researchers can control the measurement process, but the process becomes time-consuming and prone to error

Engineering Contradiction:

Improvemeasurement accuracyVSAvoidtime for wavelength optimization

Core Design Contradiction:

ReliabilityVSLoss of time

Solution Approach 1:

The system performs automatic wavelength optimization by self-determining the optimal excitation-emission wavelength pairs through computer-controlled scanning and signal-to-background ratio calculation, eliminating the need for manual intervention while maintaining measurement accuracy

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The system continuously calculates signal-to-background ratios during automated scanning and uses this feedback to identify the optimal wavelength pairs, ensuring reliable measurements through objective criteria rather than manual judgment

Inventive Principle:

Principle #23Feedback

2Measurement precision

If researchers scan across the entire wavelength range to identify wavelength ranges of interest, then they can discover optimal wavelengths, but the process becomes time-consuming

Engineering Contradiction:

Improvewavelength identification accuracyVSAvoidspeed of wavelength optimization

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The patent replaces manual mechanical scanning and visual inspection with automated computer-controlled wavelength scanning and algorithmic signal-to-background ratio calculation, significantly increasing productivity while maintaining measurement precision

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The system automatically varies excitation and emission wavelength parameters across the specified range and calculates signal-to-background ratios at each combination, efficiently identifying optimal parameters without manual intervention

Inventive Principle:

Principle #35Parameter changes

3Illumination intensity

If excitation and emission bandpasses are set close together to maximize intensity, then raw intensity is maximized, but crosstalk occurs from excitation light leaking into the emission channel

Engineering Contradiction:

Improvefluorescence intensityVSAvoidcrosstalk

Core Design Contradiction:

Illumination intensityVSObject-generated harmful factors

Solution Approach 1:

The system calculates signal-to-background ratios that account for background signal levels, providing feedback that helps identify wavelength pairs that maximize fluorescence intensity while minimizing crosstalk effects through objective quantitative criteria

Inventive Principle:

Principle #23Feedback

Solution Approach 2:

The patent introduces the signal-to-background ratio as an intermediary metric that mediates between the competing requirements of maximizing fluorescence intensity and minimizing crosstalk, allowing objective selection of optimal wavelength pairs

Inventive Principle:

Principle #24Intermediary (Mediator)

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 significantly reduces the time and error in determining the optimal wavelength pair, ensuring maximum sensitivity by automatically identifying the excitation-emission wavelength pair that yields the highest signal-to-background ratio, thus enhancing the accuracy and efficiency of spectrofluorometry.

Implementation Method 1

Fluorophores are molecular components that cause molecular fluorescence. Fluorophores absorb light energy having one wavelength and emit light energy at a different wavelength.

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Data Source

PatentUS8723139B2System and method for automatically determining optimal excitation and emission wavelengths of a fluorophore

Publication Date: 2014.05.13 MOLECULAR DEVICES LLC

AI summary

A system for performing spectrofluorometry of a fluorophore sample is provided. The system includes an input module that receives user input corresponding to spectrofluorometer settings. A control module transmits control signals for controlling the spectrofluorometer during respective wavelength scans of a fluorophore sample and a blank sample. The control signals provide for automatic execution of the wavelength scans over an excitation wavelength range and an emission wavelength range. A signal-to-background determination module automatically determines multiple signal-to-background ratios based on fluorescence measurements of the fluorophore sample and the blank sample received from the spectrofluorometer. A signal-to-background analysis module automatically determines the maximum signal-to-background ratio from the multiple signal-to-background ratios.