Total internal reflection plasmonic scattering-based fluorescence-free nanoimmunosensor probe for ultra-sensitive detection of cancer antigen 125

Chakkarapani, Suresh Kumar; Zhang, Peng; Ahn, Su−Jin; Kang, Seong Ho

doi:10.1016/j.bios.2016.01.094

Cited by 29 publications

(13 citation statements)

References 38 publications

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“…A lab-made dove prism-type TIRF setup, resembling the apparatus previously used in our group, but with some modifications, was employed (Additional file 1 : Fig. S2) [ 38 ]. The setup consisted of an Olympus BX53 upright microscope (Olympus Optical Co., Ltd., Tokyo, Japan) equipped with a × 100-oil type objective lens with numerical aperture 1.4 (Olympus, UPlanSApo) and a DIC slider with Nomarski prism (Olympus, U-DICT).…”

Section: Methodsmentioning

confidence: 99%

Quantifying intracellular trafficking of silica-coated magnetic nanoparticles in live single cells by site-specific direct stochastic optical reconstruction microscopy

et al. 2021

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Background Nanoparticles have been used for biomedical applications, including drug delivery, diagnosis, and imaging based on their unique properties derived from small size and large surface-to-volume ratio. However, concerns regarding unexpected toxicity due to the localization of nanoparticles in the cells are growing. Herein, we quantified the number of cell-internalized nanoparticles and monitored their cellular localization, which are critical factors for biomedical applications of nanoparticles. Methods This study investigates the intracellular trafficking of silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate dye [MNPs@SiO2(RITC)] in various live single cells, such as HEK293, NIH3T3, and RAW 264.7 cells, using site-specific direct stochastic optical reconstruction microscopy (dSTORM). The time-dependent subdiffraction-limit spatial resolution of the dSTORM method allowed intracellular site-specific quantification and tracking of MNPs@SiO2(RITC). Results The MNPs@SiO2(RITC) were observed to be highly internalized in RAW 264.7 cells, compared to the HEK293 and NIH3T3 cells undergoing single-particle analysis. In addition, MNPs@SiO2(RITC) were internalized within the nuclei of RAW 264.7 and HEK293 cells but were not detected in the nuclei of NIH3T3 cells. Moreover, because of the treatment of the MNPs@SiO2(RITC), more micronuclei were detected in RAW 264.7 cells than in other cells. Conclusion The sensitive and quantitative evaluations of MNPs@SiO2(RITC) at specific sites in three different cells using a combination of dSTORM, transcriptomics, and molecular biology were performed. These findings highlight the quantitative differences in the uptake efficiency of MNPs@SiO2(RITC) and ultra-sensitivity, varying according to the cell types as ascertained by subdiffraction-limit super-resolution microscopy. Graphical Abstract

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Section: Methodsmentioning

confidence: 99%

Quantifying intracellular trafficking of silica-coated magnetic nanoparticles in live single cells by site-specific direct stochastic optical reconstruction microscopy

et al. 2021

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“…The conjugation of the secondary antibody onto 80 nm AgNPs followed a previous method . The NP was functionalized by adding 30 mM MCH and 10 mM MUA dissolved in ethanol to the 80 nm AgNP solution.…”

Section: Methodsmentioning

confidence: 99%

One-Shot Dual-Code Immunotargeting for Ultra-Sensitive Tumor Necrosis Factor-α Nanosensors by 3D Enhanced Dark-Field Super-Resolution Microscopy

Lee

Chakkarapani

et al. 2018

Anal. Chem.

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Tumor necrosis factor-α (TNF-α) is a significant mediator of autoimmune diseases and an inflammatory protein biomarker. A novel method for the immunotargeting of TNF-α has been developed using three-dimensional (3D) enhanced dark-field super-resolution microscopy (3D EDF-SRM) based on ultrasensitive dual-code plasmonic nanosensing. Dual-code EDF-based 3D SRM improved the localization precision and sensitivity with a least-cubic algorithm, which provides accurate position information for the immunotargeted site. A dual-view device and digital single-lens reflex (DSLR) camera were used for simultaneous dual confirmable quantitative and qualitative immunoscreening based on enhanced dark-field scattering images. Two different sizes of silver nanoparticles (40- and 80-nm AgNPs) were compared to enhance the scattering signal of the immunotargeted plasmonic nanoprobe for the 3D EDF-SRM system. The standard TNF-α was immunotargeted at a single-molecule level and was quantitatively analyzed by measuring the scattering signals of 80 nm AgNPs on an array chip with gold-nanostages (GNSs) with 100 nm spot diameters. The localization precision in the 80 nm AgNP immunotag on the GNS narrowed to ∼9.5 nm after applying the least-cubic algorithm. The developed nanosensor exhibited a detection limit of 65 zM (1.14 ag/mL; S/N = 3) with a wide dynamic detection range of 65 zM-2.08 pM (1.14 ag/mL-36.4 pg/mL; R = 0.9921). These values are 20-33 400 000 times lower than detection limits obtained using previous methods. In addition, a recovery greater than 98% was achieved by spiking standard TNF-α into human serum samples. This method should facilitate simultaneous improvements in immunotargeting precision and ultrahigh sensitive detection of various disease-related target protein molecules at a single-molecule level.

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“…The system setup was a modification of a previously reported setup. 29 Briefly, the setup of the detection system was based on an upright Olympus BX53 microscope (Olympus). Two 671 nm, solid-state, continuous wave lasers (50 mW, SDL-671-040T, Shanghai Dream Lasers Technology Co., Ltd., Shanghai, China) for detection of gold nanoparticles (GNPs) and a 532 nm, solid-state, continuous wave laser (70 mW, SDL-532-LM-100T Shanghai Dream Lasers Technology Co., Ltd.) were the light sources for the fluorescence detection of capsaicinoids.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Plasmon-Amplified Endogenous Fluorescence Nanospectroscopic Sensor Based on Inherent Elastic Scattering for Ultratrace Ratiometric Detection of Capsaicinoids

Chakkarapani

Lee

Park³

et al. 2019

ACS Sens.

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Endogenous fluorescence imaging techniques are key for modern single-molecule quantification without the use of additional labeling probes. However, the drawback of weak fluorescence signal is the primary challenge in meeting the ever-increasing demands of single-molecule detection.Here, we report a simple and reliable method that provides up to ∼100-fold uniform fluorescence enhancement of endogenous fluorescence of the capsaicinoid molecule. The method is based on a single nanoparticle plasmon-amplified endogenous fluorescence nanospectroscopic sensor (PAEFS). This work demonstrated the applicability of PAEFS in refining sensitivity at the single-molecule level by showing ultralow limits of detection (10 6 times lower than previous reports) of fluorescence-based capsaicinoids with a wide range of linear response (18 zM to 85 pM). Spectrally overlapped capsaicinoid analogues were quantified ratiometrically to detect the analogue percentages in real samples. The novel endogenous fluorescence enhancement approach presented here represents a universal sensor for enhanced detection of single molecules using existing techniques without altering the original molecular features or using add-on labeling probes.

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Total internal reflection plasmonic scattering-based fluorescence-free nanoimmunosensor probe for ultra-sensitive detection of cancer antigen 125

Cited by 29 publications

References 38 publications

Quantifying intracellular trafficking of silica-coated magnetic nanoparticles in live single cells by site-specific direct stochastic optical reconstruction microscopy

Quantifying intracellular trafficking of silica-coated magnetic nanoparticles in live single cells by site-specific direct stochastic optical reconstruction microscopy

One-Shot Dual-Code Immunotargeting for Ultra-Sensitive Tumor Necrosis Factor-α Nanosensors by 3D Enhanced Dark-Field Super-Resolution Microscopy

Plasmon-Amplified Endogenous Fluorescence Nanospectroscopic Sensor Based on Inherent Elastic Scattering for Ultratrace Ratiometric Detection of Capsaicinoids

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