Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells
Recent studies have shown that mitochondrial morphology can modulate organelle function and greatly affect stem cell behavior, thus affecting tissue homeostasis. As such, we previously showed that the accumulation of fragmented mitochondria in aged Drosophila ovarian germline stem cells (GSCs) contr...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-12-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.13506 |
| _version_ | 1811210598955876352 |
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| author | Chi‐Hung Lin Tzu‐Yang Lin Shao‐Chun Hsu Hwei‐Jan Hsu |
| author_facet | Chi‐Hung Lin Tzu‐Yang Lin Shao‐Chun Hsu Hwei‐Jan Hsu |
| author_sort | Chi‐Hung Lin |
| collection | DOAJ |
| description | Recent studies have shown that mitochondrial morphology can modulate organelle function and greatly affect stem cell behavior, thus affecting tissue homeostasis. As such, we previously showed that the accumulation of fragmented mitochondria in aged Drosophila ovarian germline stem cells (GSCs) contributes to age‐dependent GSC loss. However, standard immunofluorescence methods to examine mitochondrial morphology yield images with insufficient resolution for rigorous analysis, while 3‐dimensional electron microscopy examination of mitochondrial morphology is labor intensive and allows only limited sampling of mitochondria. To overcome these issues, we utilized the expansion microscopy technique to expand GSC samples by 4‐fold in combination with mitochondrial immunofluorescence labeling. Here, we present a simple, inexpensive method for nanoscale optical imaging of mitochondria in the germline. This protocol may be beneficial for studies that require visualization of mitochondria or other fine subcellular structures in the Drosophila ovary. |
| first_indexed | 2024-04-12T04:57:28Z |
| format | Article |
| id | doaj.art-e72cef34e31c415fa0d107633d150f35 |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-12T04:57:28Z |
| publishDate | 2022-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-e72cef34e31c415fa0d107633d150f352022-12-22T03:47:04ZengWileyFEBS Open Bio2211-54632022-12-0112122102211010.1002/2211-5463.13506Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cellsChi‐Hung Lin0Tzu‐Yang Lin1Shao‐Chun Hsu2Hwei‐Jan Hsu3Molecular and Cell Biology, Taiwan International Graduate Program Academia Sinica Taipei TaiwanInstitute of Cellular and Organismic Biology Academia Sinica Taipei TaiwanBranch Office of Research and Development National Taiwan University College of Medicine Taipei TaiwanMolecular and Cell Biology, Taiwan International Graduate Program Academia Sinica Taipei TaiwanRecent studies have shown that mitochondrial morphology can modulate organelle function and greatly affect stem cell behavior, thus affecting tissue homeostasis. As such, we previously showed that the accumulation of fragmented mitochondria in aged Drosophila ovarian germline stem cells (GSCs) contributes to age‐dependent GSC loss. However, standard immunofluorescence methods to examine mitochondrial morphology yield images with insufficient resolution for rigorous analysis, while 3‐dimensional electron microscopy examination of mitochondrial morphology is labor intensive and allows only limited sampling of mitochondria. To overcome these issues, we utilized the expansion microscopy technique to expand GSC samples by 4‐fold in combination with mitochondrial immunofluorescence labeling. Here, we present a simple, inexpensive method for nanoscale optical imaging of mitochondria in the germline. This protocol may be beneficial for studies that require visualization of mitochondria or other fine subcellular structures in the Drosophila ovary.https://doi.org/10.1002/2211-5463.13506DrosophilaExMexpansion microscopyGSCmitochondriaovary |
| spellingShingle | Chi‐Hung Lin Tzu‐Yang Lin Shao‐Chun Hsu Hwei‐Jan Hsu Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells FEBS Open Bio Drosophila ExM expansion microscopy GSC mitochondria ovary |
| title | Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells |
| title_full | Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells |
| title_fullStr | Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells |
| title_full_unstemmed | Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells |
| title_short | Expansion Microscopy‐based imaging for visualization of mitochondria in Drosophila ovarian germline stem cells |
| title_sort | expansion microscopy based imaging for visualization of mitochondria in drosophila ovarian germline stem cells |
| topic | Drosophila ExM expansion microscopy GSC mitochondria ovary |
| url | https://doi.org/10.1002/2211-5463.13506 |
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