Inverted Microscope Liquid Column Formation for 3D Cell Observation
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Solution Overview
Problem
Existing microscope technologies are inadequate for observing 3D cultured cells like spheroids or organoids with high resolution, as they lack immersion objectives with sufficient working distance and automated liquid injection systems capable of forming liquid columns between the objective and sample containers, limiting the observation of thicker samples.
Innovation Solution
An inverted microscope system with a retaining unit, immersion objective, alignment unit, and liquid injecting unit that allows for the formation of a liquid column by injecting liquid between the bottom member and the immersion objective, enabling precise alignment and focal adjustment for observing thicker samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If an immersion objective with large NA is used to achieve high-resolution observation, then measurement precision is improved, but the working distance becomes insufficient for observing thick 3D cultured cells
Solution Approach 1:
The patent introduces a liquid column as an intermediary medium between the immersion objective and the sample container bottom. This liquid column extends the effective working distance, allowing the immersion objective to focus on deep structures within thick 3D cultured cells while maintaining optical contact through the liquid medium, thereby resolving the contradiction between high NA requirement and sufficient working distance
Solution Approach 2:
The patent transitions from a traditional planar immersion configuration to a three-dimensional liquid column configuration. By forming a vertical liquid column that extends from the sample container bottom up to the immersion objective, the system creates an additional dimensional space for light transmission, enabling both high-resolution imaging and adequate working distance for thick samples
2Adaptability or versatility
If the distance between the immersion objective and sample container is increased to accommodate thick samples, then adaptability is improved, but liquid retention becomes difficult
Solution Approach 1:
The patent replaces mechanical liquid retention methods (such as physical seals or pressure systems) with surface tension-based retention. By utilizing the natural surface tension properties of the liquid at the interface between the immersion objective and sample container, the system reliably maintains the liquid column over extended distances, enabling adaptability to thick samples without compromising liquid retention
Solution Approach 2:
The patent changes the physical parameters of the liquid interface by optimizing surface tension characteristics. Through careful selection of immersion liquid and control of environmental conditions, the system enhances surface tension effects that enable the liquid column to be retained at greater distances between the objective and sample container, thus improving both adaptability and reliability
3Extent of automation
If automated liquid injection is implemented to enable screening, then extent of automation is improved, but device complexity increases
Solution Approach 1:
The patent designs the liquid injection mechanism to serve multiple functions: it not only injects liquid to form the column but also maintains liquid levels during observation, enables sample screening, and supports focal adjustment. This multi-functionality reduces the need for separate dedicated components for each operation, thereby achieving high automation without proportionally increasing device complexity
Solution Approach 2:
The patent merges the liquid injection, liquid retention, and focal adjustment functions into a single integrated system. By combining these operations that were traditionally performed by separate mechanisms, the system achieves comprehensive automation while minimizing the increase in device complexity through functional consolidation
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-resolution observation of 3D cultured cells by maintaining a stable liquid column, increasing the retaining force and allowing for sufficient liquid volume between the objective and sample, thus overcoming the limitations of existing technologies in observing thicker samples.
Implementation Method 1
a droplet forming step of, in a state where a bottom member that retains a sample thereabove and that is optically transparent and an immersion objective that is disposed facing vertically upward with the distal end thereof facing the bottom face of the bottom member are disposed with a gap therebetween greater than the height of a droplet that can be formed on the distal end of the immersion objective, injecting liquid onto the distal end to form the droplet
Implementation Method 2
a liquid-column forming step of relatively bringing the bottom member and the immersion objective closer to each other in an optical-axis direction extending along the optical axis of the immersion objective to bring the droplet formed on the distal end into contact with the bottom face of the bottom member, thereby forming a liquid column constituted of the liquid being in contact with the distal end and the bottom face
Data Source
AI summary
An inverted microscope includes a stage supporting a bottom member; an immersion objective disposed facing vertically upward such that a distal end thereof faces a bottom face of the bottom member; an alignment unit moving the stage or the immersion objective in an optical-axis direction of the immersion objective; a liquid injecting unit injecting liquid between the bottom face and the distal end; and a controller controlling the liquid injecting unit, in a state where the bottom face and the distal end are disposed with a gap therebetween, to inject the liquid onto the distal end to form a droplet, and controlling the alignment unit to bring the bottom member and the immersion objective closer to each other to bring the droplet into contact with the bottom face, thereby forming a liquid column, and moving the stage and the immersion objective relative to each other while maintaining the liquid column.