Multispectral Imaging of Living Cell Conjugates

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Solution Overview

Problem

Current methods for studying inter-cellular communication in immune cells are limited by the need for fixed cells, which disrupt biochemical signaling, and the inability to analyze rare primary T cell and antigen presenting cell conjugates in suspension, making it difficult to model in vivo behavior effectively.

Innovation Solution

A method involving a multispectral imaging system that allows simultaneous acquisition of multiple images of living biological cells in suspension, processing these images to distinguish between individual cells and conjugates, and analyzing the synapse between cells to quantify molecular distribution changes, using techniques like image masking and signal normalization.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If cells are fixed for imaging, then image quality and stability are improved, but biochemical signaling mechanisms are disrupted

Engineering Contradiction:
Improveimage stabilityVSAvoidbiochemical signaling
Core Design Contradiction:
ReliabilityVSLoss of information

Solution Approach 1:

The patent changes the fundamental parameter of cell state from fixed to living, enabling the imaging of dynamic biochemical processes while maintaining image quality through advanced imaging techniques and computational methods that can capture and stabilize images of living cells

Inventive Principle:
Principle #35Parameter changes

2Productivity

If multiple images of conjugated cells are acquired simultaneously, then throughput is improved, but image processing complexity increases

Engineering Contradiction:
Improveimaging throughputVSAvoidimage processing
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent segments the image processing workflow into distinct modules: image acquisition, cell detection, conjugate identification, and synapse analysis. This modular approach enables simultaneous processing of multiple images while managing complexity through systematic decomposition of the overall task

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent uses template matching and reference images to simplify the analysis of multiple conjugated cell images. By creating representative templates from sample images, the system can efficiently process numerous images through pattern recognition rather than exhaustive analysis of each individual image

Inventive Principle:
Principle #26Copying

3Reliability

If cells are imaged in fluid suspension, then in vivo behavior is preserved, but cell adhesion to imaging surface is reduced

Engineering Contradiction:
Improvein vivo modelingVSAvoidcell positioning
Core Design Contradiction:
ReliabilityVSManufacturing precision

Solution Approach 1:

The patent introduces a flow cell chamber as an intermediary system that allows cells to remain in fluid suspension while passing through the imaging field. This mediator enables both in vivo-like conditions and precise imaging by controlling fluid flow to position cells optimally during the imaging process

Inventive Principle:
Principle #24Intermediary (Mediator)

4Reliability

If primary T cell and antigen presenting cell conjugates are analyzed, then physiological relevance is improved, but sample availability is limited

Engineering Contradiction:
Improvephysiological relevanceVSAvoidsample throughput
Core Design Contradiction:
ReliabilityVSQuantity of substance

Solution Approach 1:

The patent performs preliminary enrichment and isolation of primary T cell and antigen presenting cell conjugates before imaging. By preparing and concentrating the rare conjugates in advance, the system maximizes the use of limited samples and enables detailed analysis of physiologically relevant cell interactions

Inventive Principle:
Principle #10Preliminary action

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

Enables the detection and quantification of cell conjugates in flow, facilitating the study of inter-cellular communication and molecular redistribution in living cells, overcoming the limitations of standard microscopy and flow cytometry.

Implementation Method 1

a multispectral imaging system that allows simultaneous acquisition of multiple images of living biological cells in suspension

Methodology Applied
Scientific EffectLight scattering: Scattering

Implementation Method 2

using the imaging system to image the population of living biological cells, such that for each individual cell and for each cell conjugate in the population, a plurality of images of the cell or cell conjugate are simultaneously acquired

Methodology Applied
Scientific EffectOptical focusing: Focusing

Data Source

PatentEP1886139B1Methods for analyzing inter-cellular phenomena
Publication Date: 2011.08.17 AMNIS CORP
  • EP1886139B1 patent drawingFigure 1
  • EP1886139B1 patent drawingFigure 2
  • EP1886139B1 patent drawingFigure 3

AI summary

Aspects of the present invention encompass the collection of multispectral images from a population of objects, and the analysis of the collected images to measure at least one characteristic of the population, using photometric and/or morphometric features identifiable in the collection of images. In an exemplary application, the objects are biological cells. In a particularly preferred, but not limiting implementation, the plurality of images for each individual object are collected simultaneously. In an empirical study, the characteristic being measured involves the synapse between conjugated cells. The conjugated cells may represent a subpopulation of the overall population of objects that were imaged. In a particularly preferred, yet not limiting embodiment, the present invention enables the quantization of the redistribution of cellular molecules due to the conjugation of different biological cells. Significantly, such quantization is not feasible with standard microscopy and flow cytometry.