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Super-Resolution Imaging of Self-Assembled Nanocarriers Using Quantitative Spectroscopic Analysis for Cluster Extraction
journal contribution
posted on 2020-02-25, 20:47 authored by Janel
L. Davis, Yang Zhang, Sijia Yi, Fanfan Du, Ki-Hee Song, Evan A. Scott, Cheng Sun, Hao F. ZhangSelf-assembled
nanocarriers have inspired a range of applications
for bioimaging, diagnostics, and drug delivery. The noninvasive visualization
and characterization of these nanocarriers are important to understand
their structure to function relationship. However, the quantitative
visualization of nanocarriers in the sample’s native environment
remains challenging with the use of existing technologies. Single-molecule
localization microscopy (SMLM) has the potential to provide both high-resolution
visualization and quantitative analysis of nanocarriers in their native
environment. However, nonspecific binding of fluorescent probes used
in SMLM can introduce artifacts, which imposes challenges in the quantitative
analysis of SMLM images. We showed the feasibility of using spectroscopic
point accumulation for imaging in nanoscale topography (sPAINT) to
visualize self-assembled polymersomes (PS) with molecular specificity.
Furthermore, we analyzed the unique spectral signatures of Nile Red
(NR) molecules bound to the PS to reject artifacts from nonspecific
NR bindings. We further developed quantitative spectroscopic analysis
for cluster extraction (qSPACE) to increase the localization density
by 4-fold compared to sPAINT; thus, reducing variations in PS size
measurements to less than 5%. Finally, using qSPACE, we quantitatively
imaged PS at various concentrations in aqueous solutions with ∼20
nm localization precision and 97% reduction in sample misidentification
relative to conventional SMLM.
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sPAINTcluster extractionsample misidentificationnoninvasive visualizationfunction relationshiplocalization densityqSPACESingle-molecule localization microscopyQuantitative Spectroscopic AnalysisCluster Extraction Self-assembled nanocarriersSMLM imagesNR bindingsSelf-Assembled Nanocarriersnanoscale topographyartifactSuper-Resolution ImagingNile Redself-assembled polymersomesPS size measurementsspectroscopic analysisspectroscopic point accumulationdrug delivery
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