Image Cytometry via Scanned Light Sheet for 3D Cell Analysis
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Solution Overview
Problem
Conventional cytometry methods, such as flow cytometry, lack the ability to provide spatial information and subcellular distribution data, while image-based cytometry systems suffer from low throughput and require cells to be spread out in a two-dimensional layer or rely on sequential imaging.
Innovation Solution
An image cytometry system utilizing a thin sheet of light scanned across a three-dimensional suspension medium, combined with an imaging subsystem to capture reflected and emitted light, enabling three-dimensional imaging and analysis of cells without disrupting the sample, using selective plane illumination microscopy (SPIM) technology.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If flow cytometry is used for high-throughput cell analysis, then throughput and automated sample handling are improved, but spatial information and subcellular distribution data are lost
Solution Approach 1:
The patent transitions from two-dimensional flow cytometry to three-dimensional light sheet microscopy, enabling simultaneous acquisition of spatial coordinates (x, y, z) and temporal information while maintaining high throughput. The light sheet illuminates the entire sample volume, capturing cells in 3D space without sequential imaging, thus preserving spatial distribution data while achieving flow cytometry-level throughput.
2Measurement precision
If image-based cytometry is used to obtain spatial information, then spatial resolution and morphology data are improved, but throughput decreases
Solution Approach 1:
The system employs three-dimensional light sheet microscopy that illuminates the entire sample volume simultaneously, capturing all cells in the field of view in focus. This eliminates the need for z-stack acquisition and enables parallel imaging of multiple cells at high spatial resolution, achieving both morphological detail and high throughput.
Solution Approach 2:
The light sheet continuously illuminates the sample volume while cells flow through, enabling continuous image acquisition without sequential scanning. This continuous imaging approach maintains high temporal resolution and throughput while capturing spatial information, unlike traditional confocal microscopy that requires sequential z-stack acquisition.
3Measurement precision
If cells are spread in a two-dimensional layer for imaging, then focus and image quality are improved, but sample integrity and natural state are compromised
Solution Approach 1:
The patent uses three-dimensional light sheet microscopy that captures cells in their native three-dimensional suspension state without requiring them to be spread in monolayers. The light sheet illuminates the entire depth of the sample, keeping all cells in focus regardless of their position in the z-dimension, thus preserving sample integrity while maintaining image quality.
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 high-throughput, three-dimensional imaging and analysis of cells in situ, providing rich information on cell morphology and subcellular distribution without the limitations of conventional methods, allowing for repetitive readouts and tracking of biological processes.
Implementation Method 1
an imaging subsystem configured to receive light reflected, refracted, scattered and/or emitted by the cells/objects
Implementation Method 2
an imaging subsystem configured to receive light reflected, refracted, scattered and/or emitted by the cells/objects
Implementation Method 3
an imaging subsystem configured to receive light reflected, refracted, scattered and/or emitted by the cells/objects
Implementation Method 4
an imaging subsystem configured to receive light reflected, refracted, scattered and/or emitted by the cells/objects
Data Source
AI summary
The present disclosure describes image cytometry methods and systems. One such system comprises an illumination subsystem configured to generate a thin sheet of light; a scanning subsystem configured to move the sheet of light across a threedimensional suspension medium that contains cells or other objects; and an imaging subsystem configured to receive light reflected and/or emitted by the cells/objects.