Live Tissue Activity Measurement With Single-Sample Normalization
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Solution Overview
Problem
Current methods for measuring cellular activity in heterogeneous tissue samples, such as tumor biopsies, are confounded by tissue heterogeneity and require extensive dissociation or resource-intensive parallel testing, compromising the native tumor microenvironment and being impractical with scarce samples.
Innovation Solution
A single-sample assay method where a tissue sample is exposed sequentially to a control and treatment stimulus, allowing for continuous measurement of cellular activity changes over time, with internal normalization for tissue heterogeneity by calculating the slope or fold-change of secretory factors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If tissue is dissociated into smaller pieces to achieve homogeneity between treatment groups, then measurement precision is improved, but the native tumor microenvironment is lost and tissue architecture is compromised
Solution Approach 1:
The tissue sample is divided into multiple regions of interest (ROIs) within the same tissue section, allowing separate analysis of different cell populations while maintaining the overall tissue architecture and microenvironment intact. This enables homogeneous comparison without physical dissociation.
Solution Approach 2:
Multiple analysis levels are nested within the single tissue section: whole tissue analysis, regional analysis (ROIs), and cellular analysis, all performed on the same undissociated tissue sample. This nested approach allows homogeneous group comparisons while preserving the native microenvironment at each level.
2Measurement precision
If large numbers of samples are tested in parallel to compensate for heterogeneity, then measurement precision is improved, but resource requirements increase significantly
Solution Approach 1:
Multiple control and treated regions are combined within a single tissue section, allowing statistical comparison across multiple regions while using only one tissue sample. This merging approach provides the statistical power of multiple samples without the actual need for multiple scarce tissue specimens.
Solution Approach 2:
A single tissue section serves multiple functions: it contains both control and treated regions, provides multiple ROIs for analysis, and maintains the native microenvironment throughout. This multi-functional use maximizes the information obtained from limited tissue resources.
3Ease of operation
If tissue is extensively dissociated to achieve homogeneity, then ease of operation is improved, but the native tumor microenvironment and cell neighborhoods are lost
Solution Approach 1:
The tissue section is segmented into multiple regions of interest that can be independently analyzed, providing operational ease through defined analysis zones while maintaining the overall tissue architecture and native microenvironment intact.
4Productivity
If a single tissue sample is used for both control and treatment, then productivity is improved, but measurement precision may be affected by tissue heterogeneity
Solution Approach 1:
The single tissue sample is segmented into multiple distinct regions of interest, with some regions designated as control and others as treated. This segmentation allows accurate comparison between conditions while efficiently using the single tissue sample, as each ROI can be analyzed independently.
Solution Approach 2:
Control and treated regions are merged within the same tissue section, allowing direct comparison while controlling for tissue heterogeneity. The proximity of control and treated regions ensures they experience similar microenvironmental conditions, improving measurement precision.
Data Source
AI summary
Provided herein are systems and methods for measuring cellular activity within a biological sample comprising living cells over an extended time period. In particular, provided herein are techniques for measuring changes in cellular activity within a tissue sample comprising living cells (e.g., a tissue section comprising living cells) during and/or after exposure to a first stimulus (e.g., a control stimulus) at a first time point and during and/or after exposure to a second stimulus (e.g., a non-control stimulus) (e.g., a pharmaceutical stimulus) at a second time point after the first time point.