Augmented 3D super-resolution of fluorescence-free nanoparticles using enhanced dark-field illumination based on wavelength-modulation and a least-cubic algorithm.
Augmented three-dimensional (3D) subdiffraction-limited resolution of fluorescence-free single-nanoparticles was achieved with wavelength-dependent enhanced dark-field (EDF) illumination and a least-cubic algorithm. Various plasmonic nanoparticles on a glass slide (i.e., gold nanoparticles, GNPs; silver nanoparticles, SNPs; and gold nanorods, GNRs) were imaged and sliced in the z-direction to a thickness of 10 nm. Single-particle images were then compared with simulation data. The 3D coordinates of individual GNP, SNP, and GNR nanoparticles (x, y, z) were resolved by fitting the data with 3D point spread functions using a least-cubic algorithm and collation. Final, 3D super-resolution microscopy (SRM) images were obtained by resolving 3D coordinates and their Cramér-Rao lower bound-based localization precisions in an image space (530 nm × 530 nm × 300 nm) with a specific voxel size (2.5 nm × 2.5 nm × 5 nm). Compared with the commonly used least-square method, the least-cubic method was more useful for finding the center in asymmetric cases (i.e., nanorods) with high precision and accuracy. This novel 3D fluorescence-free SRM technique was successfully applied to resolve the positions of various nanoparticles on glass and gold nanospots (in vitro) as well as in a living single cell (in vivo) with subdiffraction limited resolution in 3D.
Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
Peng
P
Zhang
Department of Applied Mathematics, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
Kyungsoo
K
Kim
Department of Applied Chemistry and Institute of Natural Sciences, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
Seungah
S
Lee
Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
Suresh Kumar
SK
Chakkarapani
Department of Chemistry, Georgia State University, 308 Petit Science Center, Atlanta, GA 30303, USA.
Ning
N
Fang
Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.|Department of Applied Chemistry and Institute of Natural Sciences, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
Seong Ho
SH
Kang
D000465:Algorithms; D002460:Cell Line; D006046:Gold / Q000032:analysis; D057809:HEK293 Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D053768:Metal Nanoparticles / Q000032:analysis*; D008855:Microscopy, Electron, Scanning; D008857:Microscopy, Interference / Q000379:methods*; D012834:Silver / Q000032:analysis
D012834:Silver; D006046:Gold