3D Cell Aggregate Observation Block with Orientation Markers
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Solution Overview
Problem
Existing methods for three-dimensionally culturing cell tissues face challenges in accurately observing the three-dimensional structure and maintaining positional correspondence between the sample during culture and after sectioning, particularly when weak signals are involved, making it difficult to understand the appropriate direction of sectioning and the positional relationship between the sample before and after observation.
Innovation Solution
A block for preparation of an observation section is developed, comprising a cell aggregate with a three-dimensional shape and a continuous or intermittent marker extending in a predetermined direction, which is embedded in a polyhedral cell culture vessel, allowing for easy understanding of the sectioning direction and maintaining positional information through the use of a frame or notch as a marker.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If a block for preparation of an observation section is taken out from the vessel for sectioning, then sectioning can be performed, but it becomes difficult to understand whether the block is sectioned in an appropriate direction and to maintain positional correspondence between the sample during culture and after sectioning
Solution Approach 1:
The vessel is designed with markers (such as a frame body fitting the outer circumference and protrusions) before the sectioning process. These markers are preliminarily positioned to indicate the correct sectioning direction and maintain positional correspondence, so that when sectioning is performed later, the appropriate direction can be easily understood without losing positional information about the sample's original orientation during culture
2Ease of operation
If conventional bright-field microscopy is used to observe cell tissues, then observation can be performed, but it is difficult to recognize the three-dimensional structure of cell tissues
Solution Approach 1:
The invention transitions from conventional two-dimensional bright-field observation to three-dimensional observation by embedding the cell aggregate in a transparent block and sectioning it while maintaining spatial orientation markers. This allows observation in multiple dimensions and planes, enabling recognition of the three-dimensional structure of cell tissues that cannot be achieved with conventional single-plane bright-field microscopy
3Adaptability or versatility
If a thick culture gel layer is used for three-dimensional cell culture, then cell tissues can be cultured in three dimensions, but light does not pass through high cellular density regions making observation difficult
Solution Approach 1:
The thick culture gel layer containing the cell aggregate is segmented into multiple thinner sections through sectioning. Each section has reduced thickness that allows light to pass through high cellular density regions effectively, while collectively representing the complete three-dimensional structure. This segmentation enables observation of thick samples that would otherwise be opaque to light
Data Source
Figure 1A(i)~1B(ii)
Figure 2C(i)~2D(i)
Figure 3(i)~3(iii)
AI summary
A block for preparation of an observation section in accordance with the present disclosure includes: a cell aggregate that has a three-dimensional shape and that is embedded; and a marker that is continuous or intermittent and that extends in a predetermined direction.