Spectral imaging superlocalization microscopy for quantum dots

Shi, Xingbo; Li, Minmin; Zhao, Wenfeng; Liang, Aiye; Liu, Xiaojun; Gai, Hongwei

doi:10.1016/j.snb.2014.10.077

Cited by 8 publications

(5 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The spectral images of single QDs or single AuNPs can be obtained using a transmission grating-based spectral microscope. Detailed explanations for the spectral image have been reported in our previous publications. − Their corresponding fluorescent spectral images, which were colored in red, and their scattering spectral images, which were colored in green, are depicted in Scheme C,D, respectively. Images in Scheme C,D are merged in 2E.…”

Section: Resultsmentioning

confidence: 99%

“…The merged spectral patterns of the immunoreaction solution with antigens and the control solution without antigens are displayed in Figure A,B. The dot and the streak originating from the same single particle were paired in accordance with their positions. − The zeroth-order dot and the first-order streak from an individual AuNP were labeled with triangles in the same color. For example, at the top of Figure A, the spot and streak labeled with yellow triangles were from the same AuNP.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Multiplexed Homogeneous Immunoassay Based on Counting Single Immunocomplexes together with Dark-Field and Fluorescence Microscopy

et al. 2022

Self Cite

View full text Add to dashboard Cite

The development of multiplexed immunoassays is impeded by the difficulty in distinguishing labeled immunocomplexes from free probes and nonspecifically bound probes. Here, we attempted to overcome this issue by counting core–satellite-structured immunocomplexes simultaneously using dark-field and fluorescence microscopy. The tumor biomarkers of carcinoembryonic antigen (CEA), α-fetoprotein (AFP), and prostate-specific antigen (PSA) were chosen as model targets. Gold nanoparticles (AuNPs) with diameters of 70 nm were coated with the detection antibodies of the three targets. Quantum dot (QD) 525, QD 585, and QD 655 were modified with the capture antibodies of CEA, AFP, and PSA, respectively. Then, an immunocomplex containing one AuNP and one or several QDs was formed, whereas free and nonspecifically bound probes had either one AuNP or one QD. When observed with a transmission grating-based spectral microscope, the immunocomplexes had overlapping scattering and fluorescent spectral images and were therefore identified and quantified precisely. The biomarkers inside the immunocomplexes were recognized on the basis of the fluorescent first-order streaks of the QDs. Model biomarkers in buffer and in 12.6% blank plasma were quantified for validation. The limits of detection for CEA, PSA, and AFP in buffer were in dozens of femtomolar and were close to those in blank plasma. The results demonstrated that our approach worked well in distinguishing immunocomplexes from free and nonspecifically bound probes. The successful quantification of the three targets in five human plasma samples verified the reliability of our method in clinical applications.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Multiplexed Homogeneous Immunoassay Based on Counting Single Immunocomplexes together with Dark-Field and Fluorescence Microscopy

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Gai et al ,. inserted a transmission grating between the sample and the camera to be able to distinguish two neighboring QDs with different extent of blueing.…”

Section: Nanoparticles For Super‐resolution Fluorescence Microscopymentioning

confidence: 99%

“…Gai et al [85,86] inserted a transmission grating between the sample and the camera to be able to distinguish two neighboring QDs with different extent of blueing. As the microscope objective disperses the emitted light into zerothand first-order images, the grating generates an overlapped zeroth-order spot and two distinct first-order streaks of the two QDs.…”

Section: Semiconductor Quantum Dotsmentioning

confidence: 99%

Nanoparticle Probes for Super‐Resolution Fluorescence Microscopy

Lin

Nienhaus

2018

ChemNanoMat

View full text Add to dashboard Cite

The Abbe resolution limit of ∼200 nm of conventional light‐optical microscopy greatly restricts the observation of subcellular processes. This Abbe resolution barrier has been surpassed by super‐resolution fluorescence microscopy methods that use time as an additional dispersing dimension. Light irradiation can be employed to externally control the emission properties of the luminescent markers, or spontaneous fluctuations of the emission of individual emitters can be analyzed. Here we review the development and application of light‐emitting nanoscale particles for super‐resolution imaging and discuss their (photo)physical and (photo)chemical characteristics to offer guidance in the selection of the optimal nanoparticle marker for an intended super‐resolution imaging experiment.

show abstract

“…Multicolor intravital TPM can benefit from the use of spectral chips, composed of an array of PMTs and a prism or diffraction grating to separate the fluorescence photons (Im et al 2010; Shi et al 2012, 2015; Zimmermann et al 2014). Spectral segmentation of the wavelengths arising for the different fluorophores will be thereby both more accurate and easier to achieve.…”

Section: Introductionmentioning

confidence: 99%

Two-photon probes for in vivo multicolor microscopy of the structure and signals of brain cells

Ricard

Arroyo

et al. 2018

Brain Struct Funct

View full text Add to dashboard Cite

Imaging the brain of living laboratory animals at a microscopic scale can be achieved by two-photon microscopy thanks to the high penetrability and low phototoxicity of the excitation wavelengths used. However, knowledge of the two-photon spectral properties of the myriad fluorescent probes is generally scarce and, for many, non-existent. In addition, the use of different measurement units in published reports further hinders the design of a comprehensive imaging experiment. In this review, we compile and homogenize the two-photon spectral properties of 280 fluorescent probes. We provide practical data, including the wavelengths for optimal two-photon excitation, the peak values of two-photon action cross section or molecular brightness, and the emission ranges. Beyond the spectroscopic description of these fluorophores, we discuss their binding to biological targets. This specificity allows in vivo imaging of cells, their processes, and even organelles and other subcellular structures in the brain. In addition to probes that monitor endogenous cell metabolism, studies of healthy and diseased brain benefit from the specific binding of certain probes to pathology-specific features, ranging from amyloid-β plaques to the autofluorescence of certain antibiotics. A special focus is placed on functional in vivo imaging using two-photon probes that sense specific ions or membrane potential, and that may be combined with optogenetic actuators. Being closely linked to their use, we examine the different routes of intravital delivery of these fluorescent probes according to the target. Finally, we discuss different approaches, strategies, and prerequisites for two-photon multicolor experiments in the brains of living laboratory animals.

show abstract

Spectral imaging superlocalization microscopy for quantum dots

Cited by 8 publications

References 28 publications

Multiplexed Homogeneous Immunoassay Based on Counting Single Immunocomplexes together with Dark-Field and Fluorescence Microscopy

Multiplexed Homogeneous Immunoassay Based on Counting Single Immunocomplexes together with Dark-Field and Fluorescence Microscopy

Nanoparticle Probes for Super‐Resolution Fluorescence Microscopy

Two-photon probes for in vivo multicolor microscopy of the structure and signals of brain cells

Contact Info

Product

Resources

About