Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells
In nanomedicine, lipid‐based nanoparticles (NPs) such as liposomes (LPs) have established an important position. Precise delineation of NP interaction with cells and detailed characterization of activity are becoming essential, which mainly rely on labeling with lipophilic fluorescent molecules and...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2023-08-01
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| Series: | Small Science |
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| Online Access: | https://doi.org/10.1002/smsc.202300084 |
| _version_ | 1797739718058704896 |
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| author | Wen-Qiu Huang Peter C. Burgers Mohamadreza Amin Theo M. Luider Timo L. M. ten Hagen |
| author_facet | Wen-Qiu Huang Peter C. Burgers Mohamadreza Amin Theo M. Luider Timo L. M. ten Hagen |
| author_sort | Wen-Qiu Huang |
| collection | DOAJ |
| description | In nanomedicine, lipid‐based nanoparticles (NPs) such as liposomes (LPs) have established an important position. Precise delineation of NP interaction with cells and detailed characterization of activity are becoming essential, which mainly rely on labeling with lipophilic fluorescent molecules and assuming stable association with NPs. However, because of label separation from NPs in (biological) media, or when processed by cells, fluorescence‐based detection of an NP incorporating a single label may not necessarily indicate the actual presence of an NP but may be from the dissociated label, rendering results unreliable. Herein, flow cytometry and confocal microscopy are employed to demonstrate that to verify the localization of LPs in a cell with perfect accuracy, dual‐labeling, and contemporaneous detection of both fluorescent signals in one pixel are required. This is combined with size exclusion chromatography (SEC) and mass spectrometry measurements to indicate factors involved in label dissociation, which helps to understand the possible conditions of dissociated label and NP. It is shown that determining label colocalization with, and label dissociation from, dual‐labeled NPs are needed to provide accurate spatiotemporal insight into targeting destination (colocalized signals) and disintegration (separated signals) of NPs during intracellular processing and in studying payload delivery with precision in nanomedicine. |
| first_indexed | 2024-03-12T14:02:12Z |
| format | Article |
| id | doaj.art-d238a97915b341ba9e28f4f85692384f |
| institution | Directory Open Access Journal |
| issn | 2688-4046 |
| language | English |
| last_indexed | 2024-03-12T14:02:12Z |
| publishDate | 2023-08-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Small Science |
| spelling | doaj.art-d238a97915b341ba9e28f4f85692384f2023-08-22T05:02:14ZengWiley-VCHSmall Science2688-40462023-08-0138n/an/a10.1002/smsc.202300084Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living CellsWen-Qiu Huang0Peter C. Burgers1Mohamadreza Amin2Theo M. Luider3Timo L. M. ten Hagen4Precision Medicine in Oncology (PrMiO) Department of Pathology, and Nanomedicine Innovation Center Erasmus (NICE) Erasmus MC Cancer Institute 3015 GD Rotterdam The NetherlandsClinical and Cancer Proteomics Department of Neurology Erasmus MC 3015 GD Rotterdam The NetherlandsPrecision Medicine in Oncology (PrMiO) Department of Pathology, and Nanomedicine Innovation Center Erasmus (NICE) Erasmus MC Cancer Institute 3015 GD Rotterdam The NetherlandsClinical and Cancer Proteomics Department of Neurology Erasmus MC 3015 GD Rotterdam The NetherlandsPrecision Medicine in Oncology (PrMiO) Department of Pathology, and Nanomedicine Innovation Center Erasmus (NICE) Erasmus MC Cancer Institute 3015 GD Rotterdam The NetherlandsIn nanomedicine, lipid‐based nanoparticles (NPs) such as liposomes (LPs) have established an important position. Precise delineation of NP interaction with cells and detailed characterization of activity are becoming essential, which mainly rely on labeling with lipophilic fluorescent molecules and assuming stable association with NPs. However, because of label separation from NPs in (biological) media, or when processed by cells, fluorescence‐based detection of an NP incorporating a single label may not necessarily indicate the actual presence of an NP but may be from the dissociated label, rendering results unreliable. Herein, flow cytometry and confocal microscopy are employed to demonstrate that to verify the localization of LPs in a cell with perfect accuracy, dual‐labeling, and contemporaneous detection of both fluorescent signals in one pixel are required. This is combined with size exclusion chromatography (SEC) and mass spectrometry measurements to indicate factors involved in label dissociation, which helps to understand the possible conditions of dissociated label and NP. It is shown that determining label colocalization with, and label dissociation from, dual‐labeled NPs are needed to provide accurate spatiotemporal insight into targeting destination (colocalized signals) and disintegration (separated signals) of NPs during intracellular processing and in studying payload delivery with precision in nanomedicine.https://doi.org/10.1002/smsc.202300084high-resolution live cell imaginglipid nanoparticleslipophilic fluorescent labelsliposomesprecision nanomedicine |
| spellingShingle | Wen-Qiu Huang Peter C. Burgers Mohamadreza Amin Theo M. Luider Timo L. M. ten Hagen Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells Small Science high-resolution live cell imaging lipid nanoparticles lipophilic fluorescent labels liposomes precision nanomedicine |
| title | Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells |
| title_full | Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells |
| title_fullStr | Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells |
| title_full_unstemmed | Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells |
| title_short | Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells |
| title_sort | precision localization of lipid based nanoparticles by dual fluorescent labeling for accurate and high resolution imaging in living cells |
| topic | high-resolution live cell imaging lipid nanoparticles lipophilic fluorescent labels liposomes precision nanomedicine |
| url | https://doi.org/10.1002/smsc.202300084 |
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