Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells
Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells
The transport and targeting of internalized molecules to distinct intracellular organelles/compartments can prove challenging to visualize clearly, which can contribute to some of the difficulties associated with these studies. By combining several approaches, we show how the trafficking and processing of photoreceptor outer segments in the phagosome and autophagy-lysosomal pathways of the retinal pigment epithelium (RPE) can easily be quantified and visualized as 3D-reconstructed images. This protocol takes advantage of new developments in microscopy and image-analysis software which has the potential to help better understand dynamic intracellular processes that underlie RPE dysfunction associated with irreversible blinding diseases such as age-related macular degeneration. The method described herein can also be used to study the trafficking and co-localization of different intracellular cargos in other cell types and tissues.
3D imaging, Co-localization, Retinal pigment epithelium (RPE), Phagosome, Lysosomes, Autophagy, Confocal Microscopy
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Keeling, Eloise
3207bbdb-d391-44af-8abc-a60c08dce45b
Chatelet, David
6371fd7a-e274-4738-9ccb-3dd4dab32928
11 April 2019
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Keeling, Eloise
3207bbdb-d391-44af-8abc-a60c08dce45b
Chatelet, David
6371fd7a-e274-4738-9ccb-3dd4dab32928
Ratnayaka, J. Arjuna, Keeling, Eloise and Chatelet, David
(2019)
Study of intracellular cargo trafficking and co-localization in the phagosome and autophagy-lysosomal pathways of Retinal Pigment Epithelium (RPE) cells.
In,
Turksen, Kursad
(ed.)
Methods in Molecular Biology: Imaging and Tracking Stem Cells.
(Methods in Molecular Biology)
New York.
Humana.
(doi:10.1007/7651_2019_223).
Record type:
Book Section
Abstract
The transport and targeting of internalized molecules to distinct intracellular organelles/compartments can prove challenging to visualize clearly, which can contribute to some of the difficulties associated with these studies. By combining several approaches, we show how the trafficking and processing of photoreceptor outer segments in the phagosome and autophagy-lysosomal pathways of the retinal pigment epithelium (RPE) can easily be quantified and visualized as 3D-reconstructed images. This protocol takes advantage of new developments in microscopy and image-analysis software which has the potential to help better understand dynamic intracellular processes that underlie RPE dysfunction associated with irreversible blinding diseases such as age-related macular degeneration. The method described herein can also be used to study the trafficking and co-localization of different intracellular cargos in other cell types and tissues.
More information
Accepted/In Press date: 2019
Published date: 11 April 2019
Keywords:
3D imaging, Co-localization, Retinal pigment epithelium (RPE), Phagosome, Lysosomes, Autophagy, Confocal Microscopy
Identifiers
Local EPrints ID: 429955
URI: http://eprints.soton.ac.uk/id/eprint/429955
PURE UUID: 9d58f631-7ee9-40ee-832b-15909ee6ba21
Catalogue record
Date deposited: 15 Apr 2019 16:30
Last modified: 16 Mar 2024 04:40
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Contributors
Author:
Eloise Keeling
Author:
David Chatelet
Editor:
Kursad Turksen
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