2022 Fiscal Year Research-status Report
Establishment of a novel platform of Raman microscope for diagnosis of hydrogel-generated cancer stem cells.
| Project/Area Number |
22K18179
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| Research Institution | Hokkaido University |
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Principal Investigator |
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| Project Period (FY) |
2022-04-01 – 2024-03-31
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| Keywords | Raman imaging / Data standardadization / stemness markor / cell moprhology / molecular imaging / machine learning |
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| Outline of Annual Research Achievements |
During this fiscal year was investigated the action of PAMPS gel on myoblast cell models. It was found that Poly (2-acrylamido-2- methyl-1-propanesulphonic acid) hydrogel have a significant impact to induce an elevation of stemness markers such as Sox2, Nanog, and Oct3/4. This elevation of stemness markers have been linked to a change of cell morphology, meaning that cell that have a higher expression level tend to be more round and small whereas the one that have a low expression of stemness markers tend to be bigger and flat. This interesting fact showed that by staining
The first Raman images of the mentioned hydrogel-cell interaction system was performed with a high-speed slit-scanning Raman microscope. The quality of the measurement was performed with different metric such as signal-to-noise ratio, Laplacian operator and non-homogenous line illumination quantification. As a result the measurement protocol for this complex measurement provide high-quality data.
To exploit Hyperspectral Raman images on hydrogel substrate, two algorithms that allow to compare Data from different Raman devices and different dates of measurement was finalized. One algorithm is for single cell measurement. Another one is for 3D colony formation measurement.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
The measurement of Raman image with hydrogel-cell substrate being slightly more complex than standard substrate measurement, a slight delay can be observed compared to what was originally planned.
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| Strategy for Future Research Activity |
The hydrogel substrate is a fascinating platform for investigating the complexity of live cells, and can ultimately lead to a better understanding of cancer cell behavior when subjected to varying mechanical properties, as hydrogels can be precisely tuned.
In this fiscal year, by tuning the mechanical properties of hydrogels, and combining Raman measurement with the two algorithms developed in the previous fiscal year for analyzing such data, I will elucidate the effect of mechanical constraints on the metabolic state of myoblast cell lines and cancer cell lines. This will provide a better understanding of the origin of the elevated cancer stemness markers observed on such substrates.
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| Causes of Carryover |
Raman experiment on hydrogel being more complex than expected I decided to buy the necessary material next fiscal year when my experimental workload is robust.
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