Dilute Eosin Staining for Fluorescence Imaging
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Solution Overview
Problem
Conventional eosin staining protocols interfere with fluorescence emission from other stains or markers, making it difficult to obtain high-quality images and accurately analyze biological samples, as eosin dominates the signal and obscures contributions from other fluorescent stains and immunofluorescent labels.
Innovation Solution
Reducing the amount of eosin applied to biological samples significantly, using dilute eosin staining protocols with concentrations much lower than conventional methods, allows for selective localization and detection of eosin without interfering with fluorescence from other stains, enabling accurate imaging and analysis by using fluorescence imaging techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If conventional eosin staining protocols are used, then vivid color contrast and clear visualization of tissue structures are achieved, but fluorescence emission from other stains or markers is interfered with and obscured
Solution Approach 1:
The patent applies parameter changes by significantly reducing the eosin concentration from conventional levels (typically 0.1-1%) to dilute levels (0.001-0.01%). This parameter change allows the sample to maintain sufficient structural visualization while reducing eosin's harmful interference with fluorescence emission from other stains and markers, enabling multiplexed fluorescent imaging.
2Measurement precision
If high concentrations of eosin are applied, then spatial accuracy of eosin localization is maintained, but quantitative accuracy is compromised due to signal dominance and obscuration of other fluorescent signals
Solution Approach 1:
By changing the eosin concentration parameter to dilute levels (0.001-0.01%), the patent enables simultaneous measurement of both spatial distribution and quantitative intensity of multiple fluorescent signals without eosin dominance. The reduced eosin concentration prevents signal obscuration while maintaining sufficient staining for spatial localization.
Solution Approach 2:
The patent applies partial action by using only the minimum necessary amount of eosin required for spatial visualization, rather than conventional excessive concentrations. This partial staining approach provides sufficient structural information while leaving adequate fluorescence signal intensity for quantitative analysis of other markers.
3Object-generated harmful factors
If dilute eosin staining is used, then fluorescence from other stains is clearly detected, but the amount of eosin must be precisely controlled to maintain detectable fluorescence
Solution Approach 1:
The patent establishes specific parameter ranges for dilute eosin concentrations (0.001-0.01%) that balance two competing requirements: reducing interference with fluorescence emission while maintaining sufficient staining for visualization. These optimized parameters enable reliable detection of other fluorescent signals without excessive control complexity.
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 method allows for precise visualization and analysis of biological samples with reduced eosin interference, maintaining spatial and quantitative accuracy of eosin measurements while enabling clear detection of other fluorescent stains and markers, improving image quality and analysis accuracy.
Implementation Method 1
directing excitation light to the sample to cause each of the stains to emit fluorescence
Data Source
AI summary
Methods and systems for imaging a biological sample are disclosed, and include: (a) staining the sample with a first stain that includes eosin and at least one additional stain; (b) directing excitation light to the sample to cause each of the stains to emit fluorescence; and (c) recording images of the fluorescence emitted from the stains in the sample, where the amount of eosin in the sample is sufficiently dilute to cause the sample to have an average optical density of less than 0.10 at green wavelengths.