Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells
A key challenge in nanomedicine stems from the continued need for a systematic understanding of the delivery of nanoparticles in live cells. Complexities in delivery are often influenced by the biophysical characteristics of nanoparticles, where even subtle changes to nanoparticle designs can alter...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/11/4/642 |
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| author | Aria Ahmed-Cox Alexander M. Macmillan Elvis Pandzic Renee M. Whan Maria Kavallaris |
| author_facet | Aria Ahmed-Cox Alexander M. Macmillan Elvis Pandzic Renee M. Whan Maria Kavallaris |
| author_sort | Aria Ahmed-Cox |
| collection | DOAJ |
| description | A key challenge in nanomedicine stems from the continued need for a systematic understanding of the delivery of nanoparticles in live cells. Complexities in delivery are often influenced by the biophysical characteristics of nanoparticles, where even subtle changes to nanoparticle designs can alter cellular uptake, transport and activity. Close examination of these processes, especially with imaging, offers important insights that can aid in future nanoparticle design or translation. Rapid fluorescence lifetime imaging microscopy (RapidFLIM) is a potentially valuable technology for examining intracellular mechanisms of nanoparticle delivery by directly correlating visual data with changes in the biological environment. To date, applications for this technology in nanoparticle research have not been explored. A PicoQuant RapidFLIM system was used together with commercial silica nanoparticles to follow particle uptake in glioblastoma cells. Importantly, RapidFLIM imaging showed significantly improved image acquisition speeds over traditional FLIM, which enabled the tracking of nanoparticle uptake into subcellular compartments. We determined mean lifetime changes and used this to delineate significant changes in nanoparticle lifetimes (>0.39 ns), which showed clustering of these tracks proximal to both extracellular and nuclear membrane boundaries. These findings demonstrate the ability of RapidFLIM to track, localize and quantify changes in single nanoparticle fluorescence lifetimes and highlight RapidFLIM as a valuable tool for multiparameter visualization and analysis of nanoparticle molecular dynamics in live cells. |
| first_indexed | 2024-03-09T22:20:50Z |
| format | Article |
| id | doaj.art-14a57a8c6c0a410681e44688bdfeb25c |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T22:20:50Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-14a57a8c6c0a410681e44688bdfeb25c2023-11-23T19:14:36ZengMDPI AGCells2073-44092022-02-0111464210.3390/cells11040642Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live CellsAria Ahmed-Cox0Alexander M. Macmillan1Elvis Pandzic2Renee M. Whan3Maria Kavallaris4Children’s Cancer Institute, Lowy Cancer Research Center, UNSW Sydney, Randwick, NSW 2031, AustraliaKatharina Gaus Light Microscopy Facility, Mark Wainwright Analytical Center, UNSW Sydney, Sydney, NSW 2031, AustraliaKatharina Gaus Light Microscopy Facility, Mark Wainwright Analytical Center, UNSW Sydney, Sydney, NSW 2031, AustraliaKatharina Gaus Light Microscopy Facility, Mark Wainwright Analytical Center, UNSW Sydney, Sydney, NSW 2031, AustraliaChildren’s Cancer Institute, Lowy Cancer Research Center, UNSW Sydney, Randwick, NSW 2031, AustraliaA key challenge in nanomedicine stems from the continued need for a systematic understanding of the delivery of nanoparticles in live cells. Complexities in delivery are often influenced by the biophysical characteristics of nanoparticles, where even subtle changes to nanoparticle designs can alter cellular uptake, transport and activity. Close examination of these processes, especially with imaging, offers important insights that can aid in future nanoparticle design or translation. Rapid fluorescence lifetime imaging microscopy (RapidFLIM) is a potentially valuable technology for examining intracellular mechanisms of nanoparticle delivery by directly correlating visual data with changes in the biological environment. To date, applications for this technology in nanoparticle research have not been explored. A PicoQuant RapidFLIM system was used together with commercial silica nanoparticles to follow particle uptake in glioblastoma cells. Importantly, RapidFLIM imaging showed significantly improved image acquisition speeds over traditional FLIM, which enabled the tracking of nanoparticle uptake into subcellular compartments. We determined mean lifetime changes and used this to delineate significant changes in nanoparticle lifetimes (>0.39 ns), which showed clustering of these tracks proximal to both extracellular and nuclear membrane boundaries. These findings demonstrate the ability of RapidFLIM to track, localize and quantify changes in single nanoparticle fluorescence lifetimes and highlight RapidFLIM as a valuable tool for multiparameter visualization and analysis of nanoparticle molecular dynamics in live cells.https://www.mdpi.com/2073-4409/11/4/642fluorescent lifetimenanoparticle uptaketrackinglive cell imaging |
| spellingShingle | Aria Ahmed-Cox Alexander M. Macmillan Elvis Pandzic Renee M. Whan Maria Kavallaris Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells Cells fluorescent lifetime nanoparticle uptake tracking live cell imaging |
| title | Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells |
| title_full | Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells |
| title_fullStr | Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells |
| title_full_unstemmed | Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells |
| title_short | Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells |
| title_sort | application of rapid fluorescence lifetime imaging microscopy rapidflim to examine dynamics of nanoparticle uptake in live cells |
| topic | fluorescent lifetime nanoparticle uptake tracking live cell imaging |
| url | https://www.mdpi.com/2073-4409/11/4/642 |
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