Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager
Summary: Autophagy is an intracellular self-degradation process in which part of the cytoplasm, aggregates, or damaged organelles are degraded in lysosomes. Lysophagy is a specific form of selective autophagy responsible for clearing damaged lysosomes. Here, we present a protocol for inducing lysoso...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723001946 |
| _version_ | 1797843857018191872 |
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| author | Keisuke Tabata Marika Saeki Tamotsu Yoshimori Maho Hamasaki |
| author_facet | Keisuke Tabata Marika Saeki Tamotsu Yoshimori Maho Hamasaki |
| author_sort | Keisuke Tabata |
| collection | DOAJ |
| description | Summary: Autophagy is an intracellular self-degradation process in which part of the cytoplasm, aggregates, or damaged organelles are degraded in lysosomes. Lysophagy is a specific form of selective autophagy responsible for clearing damaged lysosomes. Here, we present a protocol for inducing lysosomal damage in cultured cells and assessing lysosomal damage using a high-content imager and software program. We describe steps for induction of lysosomal damage, image acquisition with spinning disk confocal microscopy, and image analysis using Pathfinder. We then detail data analysis of the clearance of damaged lysosomes.For complete details on the use and execution of this protocol, please refer to Teranishi et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-09T17:12:47Z |
| format | Article |
| id | doaj.art-894bba73fa71486f960726e7d419b6eb |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-09T17:12:47Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-894bba73fa71486f960726e7d419b6eb2023-04-20T04:37:46ZengElsevierSTAR Protocols2666-16672023-06-0142102236Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imagerKeisuke Tabata0Marika Saeki1Tamotsu Yoshimori2Maho Hamasaki3Laboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Bioscience, Osaka University, Osaka 565-0871, Japan; Department of Genetics, Graduate School of Medicine, Osaka University, Osaka 565-0871, JapanLaboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Bioscience, Osaka University, Osaka 565-0871, JapanLaboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Bioscience, Osaka University, Osaka 565-0871, Japan; Department of Genetics, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan; Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka 565-0871, JapanLaboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Bioscience, Osaka University, Osaka 565-0871, Japan; Department of Genetics, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan; Corresponding authorSummary: Autophagy is an intracellular self-degradation process in which part of the cytoplasm, aggregates, or damaged organelles are degraded in lysosomes. Lysophagy is a specific form of selective autophagy responsible for clearing damaged lysosomes. Here, we present a protocol for inducing lysosomal damage in cultured cells and assessing lysosomal damage using a high-content imager and software program. We describe steps for induction of lysosomal damage, image acquisition with spinning disk confocal microscopy, and image analysis using Pathfinder. We then detail data analysis of the clearance of damaged lysosomes.For complete details on the use and execution of this protocol, please refer to Teranishi et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723001946Cell BiologyCell CultureHigh-throughput ScreeningMicroscopy |
| spellingShingle | Keisuke Tabata Marika Saeki Tamotsu Yoshimori Maho Hamasaki Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager STAR Protocols Cell Biology Cell Culture High-throughput Screening Microscopy |
| title | Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager |
| title_full | Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager |
| title_fullStr | Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager |
| title_full_unstemmed | Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager |
| title_short | Monitoring and assessment of lysosomal membrane damage in cultured cells using the high-content imager |
| title_sort | monitoring and assessment of lysosomal membrane damage in cultured cells using the high content imager |
| topic | Cell Biology Cell Culture High-throughput Screening Microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723001946 |
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