Narrowband Filter Sets for ELISPOT Cytokine Detection
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Solution Overview
Problem
The existing ELISPOT methods face challenges in accurately detecting cells that secrete two or more different messengers due to the limitations of dual filters, which are transmissive for a wide spectral range, leading to reduced measurement accuracy, especially when messengers are secreted in different amounts.
Innovation Solution
The method employs narrowband filters for each dye, allowing for selective detection of immunocells that secrete two or more different cytokines by using different filter sets with narrowband excitation and blocking filters, preventing luminescence interference and enhancing sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If dual filters with wide spectral range are used for detecting cells secreting multiple messengers, then the device complexity is reduced, but the measurement precision deteriorates due to luminescence interference and superimposition
Solution Approach 1:
The patent divides the filter system into multiple separate filter sets, each optimized for a specific dye/cytokine combination. Instead of using a single dual filter for multiple messengers, the invention segments the detection into individual filter sets that can be sequentially applied, eliminating spectral overlap and luminescence interference between different cytokine detections.
Solution Approach 2:
The invention changes the spectral parameters of the filters by using narrowband filters with specific transmission ranges matched to the emission spectra of individual dyes. This parameter optimization allows precise detection of each cytokine's luminescence signal without interference from other dyes, improving measurement precision while maintaining manageable device complexity through systematic filter selection.
2Measurement precision
If narrowband filters are used for each dye in multiple filter sets, then the measurement precision improves by avoiding luminescence superimpositions, but the device complexity increases
Solution Approach 1:
The patent creates a universal filter set system where each filter set can be used with different dye combinations through systematic matching. The narrowband filters are designed with standard spectral characteristics that can be applied across multiple cytokine detection scenarios, allowing the same filter sets to serve multiple functions in different ELISPOT configurations, thereby managing device complexity through standardized multi-functional components.
3Ease of operation
If conventional dual filters are used for measuring cells secreting two or more messengers, then the ease of operation is maintained, but the reliability deteriorates due to reduced measurement accuracy
Solution Approach 1:
The invention introduces dynamic filtering by using sequentially applicable narrowband filter sets that can be switched between different detection channels. This dynamic approach allows the system to adapt to different cytokine combinations and secretion levels, improving reliability by selecting the appropriate filter set for each specific measurement scenario while maintaining ease of operation through automated or standardized filter changes.
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 increases the sensitivity for detecting immunocells that secrete multiple cytokines by avoiding luminescence superimpositions, enabling precise measurement of cells secreting at least interferon-γ and interleukin-2, aiding in distinguishing between acute and latent infections.
Implementation Method 1
visualizing the cells with the aid of at least two different dyes and at least two different filter sets, the filter sets including narrowband filters
Implementation Method 2
measuring the number of immunocompetent cells which secrete at least two different cytokines... visualizing the cells with the aid of at least two different dyes
Data Source
AI summary
An ELISPOT method of in vitro diagnosis of tuberculosis includes enriching or separating liquid blood fraction from its cellular components including visualizing immunocompetent cells by detecting different dyes and using at least two different filter sets, each including one narrowband excitation filter and one narrowband blocking filter, wherein the narrowband excitation filter is transmissive for light provided for luminescent excitation of a respective dye and the narrowband blocking filter is transmissive for light emitted by a respective dye and measuring the number of immunocompetent cells that secrete at least interferon-γ and interleukin-2 as a reaction to an antigen by superimposing at least two two-dimensional images generated during visualization to avoid luminescence of different dyes interfering and strong luminescence being superimposed on weak luminescence.