NSOM/QD-based fluorescence–topographic image fusion directly reveals nano-spatial peak–valley polarities of CD69 and CD71 activation molecules on cell-membrane fluctuations during T-cell activation

Zhong, Liu; Zhang, Zhun; Lu, Xiaoxu; Huang, Dan; Chen, Crystal Y.; Wang, Thomas J.; Chen, Zheng W.

doi:10.1016/j.imlet.2011.06.003

Cited by 11 publications

(9 citation statements)

References 23 publications

(33 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As such, NSOM used in conjunction with QDs is the best technique by which the distribution of membrane receptors can be measured at the nanometer scale[30][31][32][33][34][35][36]. In this study, our results indicate that VEGF can induce directed migration of MSCs through the formation of (Focal Adhesion Kinase) FAK and cytoskeletal rearrangements.…”

mentioning

confidence: 58%

“…Some studies suggest that CD44 plays a key role in cytoskeletal rearrangement [27], takes part in a CD44-Src-integrin signaling axis in lipid rafts [28], and most importantly, CD44 can be a co-receptor of VEGFR-2 [29]. [30]. Stimulated emission depletion (STED) and photoactivatable localization microscopy (PALM) lack simultaneous topographic information [31].…”

Section: Accepted Manuscriptmentioning

confidence: 99%

“…Dual-color immunofluorescence labeling for mAbs and QD conjugates was performed according to the methods described by Chen et al [30][31] …”

Section: Dual-color Immunofluorescence Labelingmentioning

confidence: 99%

See 2 more Smart Citations

Migration mechanism of mesenchymal stem cells studied by QD/NSOM

Chen¹,

Chen²,

Guo³

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

Self Cite

View full text Add to dashboard Cite

The migration of mesenchymal stem cells (MSCs) plays a key role in tumor-targeted delivery vehicles and tumor-related stroma formation. However, there so far has been no report on the distribution of cell surface molecules during the VEGF-induced migration of MSCs. Here, we have utilized near-field scanning optical microscopy (NSOM) combined with fluorescent quantum dot (QD)-based nano-technology to capture the functional relationship between CD44 and CD29 adhesion molecules on MSCs and the effect of their spatial rearrangements. Before VEGF-induced migration of MSCs, both CD44 and CD29 formed 200-220 nm nano-domains respectively, with little co-localization between the two types of domains. Surprisingly, the size of the CD44 nano-domain rapidly increased in size to 295 nm and apparently larger aggregates were formed following MSC treatment with VEGF for 10 min, while the area of co-localization increased to 0.327 μm2. Compared with CD44, CD29 was activated obviously later, for the fact that CD29 aggregation didn't appear until 30 min after VEGF treatment. Consistently, its co-localization area increased to 0.917 μm2. The CD44 and CD29 nano-domains further aggregated into larger nano-domains or even formed micro-domains on the membrane of activated MSCs. The aggregation and co-localization of these molecules promoted FAK formation and cytoskeleton rearrangement. All of the above changes induced by VEGF contributed to MSC migration. Taken together, our data of NSOM-based dual color fluorescent imaging demonstrated for the first time that CD44, together with CD29, involved in VEGF-induced migration of MSCs through the interaction between CD44 and its co-receptor of VEGFR-2.

show abstract

mentioning

confidence: 58%

Section: Accepted Manuscriptmentioning

confidence: 99%

See 1 more Smart Citation

Migration mechanism of mesenchymal stem cells studied by QD/NSOM

Chen¹,

Chen²,

Guo³

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

Self Cite

View full text Add to dashboard Cite

show abstract

“…We have recently innovated the use of near-field scanning optical microscopy (NSOM) [ 19 , 20 ] and quantum dots (QD) based nanotechnology through dipole-polarization and dual-color to visualize nanoscale distribution and organization of antigen-specific TCR/CD3, coreceptor CD4, CD8, and nanospatial relationship between TCR/CD3 and CD4 or CD8 in sustained activation of primary T-cells [ 21 , 22 ]. In the current study, we intend to directly visualize nanoscale nanospatial relationship between TCR/CD3 and GM1 before and after T-cells activation, moreover to detect whether GM1 clustering or TCR/CD3 nanodomains are required for PKC θ signaling cascades and T-cell activation.…”

Section: Introductionmentioning

confidence: 99%

NSOM/QD-Based Visualization of GM1 Serving as Platforms for TCR/CD3 Mediated T-Cell Activation

Zhong

Zhang

et al. 2013

BioMed Research International

Self Cite

View full text Add to dashboard Cite

Direct molecular imaging of nanoscale relationship between T-cell receptor complexes (TCR/CD3) and gangliosidosis GM1 before and after T-cell activation has not been reported. In this study, we made use of our expertise of near-field scanning optical microscopy(NSOM)/immune-labeling quantum dots- (QD-)based dual-color imaging system to visualize nanoscale profiles for distribution and organization of TCR/CD3, GM1, as well as their nanospatial relationship and their correlation with PKCθ signaling cascade during T-cell activation. Interestingly, after anti-CD3/anti-CD28 Ab co-stimulation, both TCR/CD3 and GM1 were clustered to form nanodomains; moreover, all of TCR/CD3 nanodomains were colocalized with GM1 nanodomains, indicating that the formation of GM1 nanodomains was greatly correlated with TCR/CD3 mediated signaling. Specially, while T-cells were pretreated with PKCθ signaling inhibitor rottlerin to suppress IL-2 cytokine production, no visible TCR/CD3 nanodomains appeared while a lot of GM1 nanodomains were still observed. However, while T-cells are pretreated with PKCα β signaling inhibitor GÖ6976 to suppress calcium-dependent manner, all of TCR/CD3 nanodomains were still colocalized with GM1 nanodomains. These findings possibly support the notion that the formation of GM1 nanodomains indeed serves as platforms for the recruitment of TCR/CD3 nanodomains, and TCR/CD3 nanodomains are required for PKCθ signaling cascades and T-cell activation

show abstract

“…These methods include flow cytometry (also known as fluorescence-activated cell sorting), [14][15][16][17] electron microscopy (EM), [18][19][20] and so-called super-resolution microscopy. [21][22][23][24] For example, scanning EM (SEM) and transmission EM (TEM) are common methods for imaging NP uptake in vitro and ex vivo. Gupta et al 25 and Gupta and Gupta 26 showed the accumulation of SPIONs at the cell surface by SEM and the location of SPIONs inside single human fibroblasts by TEM.…”

Section: Introductionmentioning

confidence: 99%

Nanobarcoding: detecting nanoparticles in biological samples using in situ polymerase chain reaction

Eustaquio¹,

Leary²

2012

IJN

View full text Add to dashboard Cite

Background: Determination of the fate of nanoparticles (NPs) in a biological system, or NP biodistribution, is critical in evaluating an NP formulation for nanomedicine. Current methods to determine NP biodistribution are greatly inadequate, due to their limited detection thresholds. Herein, proof of concept of a novel method for improved NP detection based on in situ polymerase chain reaction (ISPCR), coined "nanobarcoding," is demonstrated. Methods: Nanobarcoded superparamagnetic iron oxide nanoparticles (NB-SPIONs) were characterized by dynamic light scattering, zeta potential, and hyperspectral imaging measurements. Cellular uptake of Cy5-labeled NB-SPIONs (Cy5-NB-SPIONs) was imaged by confocal microscopy. The feasibility of the nanobarcoding method was first validated by solution-phase PCR and "pseudo"-ISPCR before implementation in the model in vitro system of HeLa human cervical adenocarcinoma cells, a cell line commonly used for ISPCR-mediated detection of human papilloma virus (HPV). Results: Dynamic light-scattering measurements showed that NB conjugation stabilized SPION size in different dispersion media compared to that of its precursor, carboxylated SPIONs (COOHSPIONs), while the zeta potential became more positive after NB conjugation. Hyperspectral imaging confirmed NB conjugation and showed that the NB completely covered the SPION surface. Solution-phase PCR and pseudo-ISPCR showed that the expected amplicons were exclusively generated from the NB-SPIONs in a dose-dependent manner. Although confocal microscopy revealed minimal cellular uptake of Cy5-NB-SPIONs at 50 nM over 24 hours in individual cells, ISPCR detected definitive NB-SPION signals inside HeLa cells over large sample areas. Conclusion: Proof of concept of the nanobarcoding method has been demonstrated in in vitro systems, but the technique needs further development before its widespread use as a standardized assay.

show abstract

NSOM/QD-based fluorescence–topographic image fusion directly reveals nano-spatial peak–valley polarities of CD69 and CD71 activation molecules on cell-membrane fluctuations during T-cell activation

Cited by 11 publications

References 23 publications

Migration mechanism of mesenchymal stem cells studied by QD/NSOM

Migration mechanism of mesenchymal stem cells studied by QD/NSOM

NSOM/QD-Based Visualization of GM1 Serving as Platforms for TCR/CD3 Mediated T-Cell Activation

Nanobarcoding: detecting nanoparticles in biological samples using in situ polymerase chain reaction

Contact Info

Product

Resources

About