An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images
Abstract Background Characterizing membrane dynamics is a key issue to understand cell exchanges with the extra-cellular medium. Total internal reflection fluorescence microscopy (TIRFM) is well suited to focus on the late steps of exocytosis at the plasma membrane. However, it is still a challengin...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-07-01
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| Series: | BMC Bioinformatics |
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| Online Access: | http://link.springer.com/article/10.1186/s12859-017-1765-y |
| _version_ | 1818439566345895936 |
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| author | Antoine Basset Patrick Bouthemy Jérôme Boulanger François Waharte Jean Salamero Charles Kervrann |
| author_facet | Antoine Basset Patrick Bouthemy Jérôme Boulanger François Waharte Jean Salamero Charles Kervrann |
| author_sort | Antoine Basset |
| collection | DOAJ |
| description | Abstract Background Characterizing membrane dynamics is a key issue to understand cell exchanges with the extra-cellular medium. Total internal reflection fluorescence microscopy (TIRFM) is well suited to focus on the late steps of exocytosis at the plasma membrane. However, it is still a challenging task to quantify (lateral) diffusion and estimate local dynamics of proteins. Results A new model was introduced to represent the behavior of cargo transmembrane proteins during the vesicle fusion to the plasma membrane at the end of the exocytosis process. Two biophysical parameters, the diffusion coefficient and the release rate parameter, are automatically estimated from TIRFM image sequences, to account for both the lateral diffusion of molecules at the membrane and the continuous release of the proteins from the vesicle to the plasma membrane. Quantitative evaluation on 300 realistic computer-generated image sequences demonstrated the efficiency and accuracy of the method. The application of our method on 16 real TIRFM image sequences additionally revealed differences in the dynamic behavior of Transferrin Receptor (TfR) and Langerin proteins. Conclusion An automated method has been designed to simultaneously estimate the diffusion coefficient and the release rate for each individual vesicle fusion event at the plasma membrane in TIRFM image sequences. It can be exploited for further deciphering cell membrane dynamics. |
| first_indexed | 2024-12-14T17:58:30Z |
| format | Article |
| id | doaj.art-7c49a51eccc44918bfb2bc9654c3005d |
| institution | Directory Open Access Journal |
| issn | 1471-2105 |
| language | English |
| last_indexed | 2024-12-14T17:58:30Z |
| publishDate | 2017-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Bioinformatics |
| spelling | doaj.art-7c49a51eccc44918bfb2bc9654c3005d2022-12-21T22:52:30ZengBMCBMC Bioinformatics1471-21052017-07-0118111010.1186/s12859-017-1765-yAn extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy imagesAntoine Basset0Patrick Bouthemy1Jérôme Boulanger2François Waharte3Jean Salamero4Charles Kervrann5Inria, Campus de BeaulieuInria, Campus de BeaulieuInstitut Curie, PSL Research University, CNRS UMR 144 and PICT-Cell and Tissue Imaging FacilityInstitut Curie, PSL Research University, CNRS UMR 144 and PICT-Cell and Tissue Imaging FacilityInstitut Curie, PSL Research University, CNRS UMR 144 and PICT-Cell and Tissue Imaging FacilityInria, Campus de BeaulieuAbstract Background Characterizing membrane dynamics is a key issue to understand cell exchanges with the extra-cellular medium. Total internal reflection fluorescence microscopy (TIRFM) is well suited to focus on the late steps of exocytosis at the plasma membrane. However, it is still a challenging task to quantify (lateral) diffusion and estimate local dynamics of proteins. Results A new model was introduced to represent the behavior of cargo transmembrane proteins during the vesicle fusion to the plasma membrane at the end of the exocytosis process. Two biophysical parameters, the diffusion coefficient and the release rate parameter, are automatically estimated from TIRFM image sequences, to account for both the lateral diffusion of molecules at the membrane and the continuous release of the proteins from the vesicle to the plasma membrane. Quantitative evaluation on 300 realistic computer-generated image sequences demonstrated the efficiency and accuracy of the method. The application of our method on 16 real TIRFM image sequences additionally revealed differences in the dynamic behavior of Transferrin Receptor (TfR) and Langerin proteins. Conclusion An automated method has been designed to simultaneously estimate the diffusion coefficient and the release rate for each individual vesicle fusion event at the plasma membrane in TIRFM image sequences. It can be exploited for further deciphering cell membrane dynamics.http://link.springer.com/article/10.1186/s12859-017-1765-yTIRF microscopyVesicle fusion modelMolecule diffusionProtein release rateModel fittingExocytosis |
| spellingShingle | Antoine Basset Patrick Bouthemy Jérôme Boulanger François Waharte Jean Salamero Charles Kervrann An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images BMC Bioinformatics TIRF microscopy Vesicle fusion model Molecule diffusion Protein release rate Model fitting Exocytosis |
| title | An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images |
| title_full | An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images |
| title_fullStr | An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images |
| title_full_unstemmed | An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images |
| title_short | An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images |
| title_sort | extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from tirf microscopy images |
| topic | TIRF microscopy Vesicle fusion model Molecule diffusion Protein release rate Model fitting Exocytosis |
| url | http://link.springer.com/article/10.1186/s12859-017-1765-y |
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