A versatile microfluidic device for highly inclined thin illumination microscopy in the moss Physcomitrella patens
| Autor: | Gohta Goshima, Elena Kozgunova |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine Materials science Microtubule dynamics Cell Survival Microfluidics lcsh:Medicine Physcomitrella patens 01 natural sciences Microtubules Article Imaging 03 medical and health sciences Molecular level Microtubule Lab-On-A-Chip Devices Microscopy Cytoskeleton lcsh:Science Multidisciplinary biology Lab-on-a-chip lcsh:R biology.organism_classification Bryopsida 030104 developmental biology Germ Cells Biophysics lcsh:Q 010606 plant biology & botany Plant cytoskeleton |
| Zdroj: | Scientific Reports, Vol 9, Iss 1, Pp 1-8 (2019) Scientific Reports |
| DOI: | 10.1101/660480 |
| Popis: | High-resolution microscopy is a valuable tool for studying cellular processes, such as signalling, membrane trafficking, or cytoskeleton remodelling. Several techniques of inclined illumination microscopy allow imaging at a near single molecular level; however, the application of these methods to plant cells is limited, owing to thick cell walls as well as the necessity to excise a part of the tissue for sample preparation. In this study, we utilised a simple, easy-to-use microfluidic device for highly inclined and laminated optical sheet (HILO) microscopy using a model plant Physcomitrella patens. We demonstrated that the shallow microfluidic device can be used for long-term culture of living cells and enables high-resolution HILO imaging of microtubules without perturbing their dynamics. In addition, our microdevice allows the supply and robust washout of compounds during HILO microscopy imaging, for example, to perform a microtubule regrowth assay. Furthermore, we tested long-term (48 h) HILO imaging using a microdevice and visualised the developmental changes in the microtubule dynamics during tissue regeneration. These novel applications of the microfluidic device provide a valuable resource for studying molecular dynamics in living plant cells. |
| Databáze: | OpenAIRE |
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