A nanobiosensor for dynamic single cell analysis during microvascular self-organization

Abstract

The formation of microvascular networks plays essential roles in regenerative medicine and tissue engineering. Nevertheless, the self-organization mechanisms underlying the dynamic morphogenic process are poorly understood due to a paucity of effective tools for mapping the spatiotemporal dynamics of single cell behaviors. By establishing a single cell nanobiosensor along with live cell imaging, we perform dynamic single cell analysis of the morphology, displacement, and gene expression during microvascular self-organization. Dynamic single cell analysis reveals that endothelial cells self-organize into subpopulations with specialized phenotypes to form microvascular networks and identifies the involvement of Notch1-Dll4 signaling in regulating the cell subpopulations. The cell phenotype correlates with the initial Dll4 mRNA expression level and each subpopulation displays a unique dynamic Dll4 mRNA expression profile. Pharmacological perturbations and RNA interference of Notch1-Dll4 signaling modulate the cell subpopulations and modify the morphology of the microvascular network. Taken together, the nanobiosensor enables a dynamic single cell analysis approach underscoring the importance of Notch1-Dll4 signaling in microvascular self-organization.

Linseed hydrogel-mediated green synthesis of silver nanoparticles for antimicrobial and wound-dressing applications

“…1 Unique electronic, 2 catalytic, 3 and optical 4 properties of such metal NPs have led to their potential applications in bioconjugation, biosensing, 5,6 and surface-enhanced Raman scattering. 7 Although NPs have been synthesized from a large variety of metals, silver nanoparticles (Ag NPs) have gained special importance over the last few years.…”

Intercellular Tension Negatively Regulates Angiogenic Sprouting of Endothelial Tip Cells via Notch1‐Dll4 Signaling

“…In contrast, nocodazole treatment downregulated the expression for Notch1 (22.5±4.4%) and Dll4 (30.7±8.9%). The negative effects of mechanical force on Notch1-Dll4 signaling were also confirmed at the transcription level using a live cell biosensor (Figure 5C and supplementary Figure S6) [13] . In particular, gold nanorod based single cell biosensors with specific locked nucleic acid probes were applied to detect the expressions of Notch1 and Dll4 mRNA.…”

“…The laser ablation experiment was performed by focusing the near infrared laser to a desired location of the network for less than two seconds (Figure 6C). The photothermal effect was previously optimized for single cell ablation [13a] . Retraction of the cells was observed immediately after ablation of the network, indicating that the cells were under intercellular tension.…”

“…Analysis of single cell gene expression was performed using gold nanorods (GNR) with locked nucleic acid (LNA) probes targeting specific mRNA sequences [13a, 13b] . Mercaptoundecyltrimethylammonnium bromide coated GNR with 10 nm diameter and 67 nm length (Nanopartz) were used.…”

A gapmer aptamer nanobiosensor for real-time monitoring of transcription and translation in single cells

“…To address the need for high-throughput single cell analysis, we developed a nanobiosensor for intracellular detection of mRNA expression in live cells and tissues 16–18 . The nanobiosensor consists of locked nucleic acid (LNA) probes and gold nanorods (GNRs).…”