Probing Membrane Transport of Single Live Cells Using Single-Molecule Detection and Single Nanoparticle Assay

2007 ◽  
pp. 41-70 ◽  
Author(s):  
Xiao-Hong Nancy Xu ◽  
Yujun Song ◽  
Prakash Nallathamby
Keyword(s):  
Membrane Transport ◽  
Single Molecule ◽  
Single Molecule Detection ◽  
Live Cells ◽  
Single Nanoparticle

Synthetic Glass Nanopore for Single Molecule Detection

2012 ◽  
Vol 229-231 ◽  
pp. 197-200
Author(s):  
Xiu Hua Sun ◽  
Chang Lu Gao ◽  
Li Qun Gu
Keyword(s):  
Pore Structure ◽  
Single Molecule ◽  
Second Messengers ◽  
Single Molecule Detection ◽  
Target Species ◽  
Single Nanoparticle ◽  
Molecular Scale ◽  
Drug Compounds ◽  
Target Molecules ◽  
Nanopore Sensors

The molecular-scale pore structure, called nanopore, interacting with target molecules in its functionalized lumen, can produce characteristic changes in the pore conductance, which allows us to identify single molecules and simultaneously quantify each target species in the mixture. Nanopore sensors have been created for tremendous biomedical detections, with targets ranging from metal ions, drug compounds and cellular second messengers, to proteins and DNAs. Here we will review our recent discoveries with a lab-in-hand glass nanopore: single-molecule discrimination of chiral enantiomers with a trapped cyclodextrin, sensing of bioterrorist agent ricin and site-directed capturing a single nanoparticle.

scholarly journals Single-molecule detection: Unravelling surface receptor diffusion in live neurons

2005 ◽  
Vol 27 (5) ◽  
pp. 5-8 ◽  
Author(s):  
Laurent Groc ◽  
Daniel Choquet ◽  
Brahim Lounis ◽  
Laurent Cognet
Keyword(s):  
Single Molecule ◽  
Cellular Imaging ◽  
Single Molecule Detection ◽  
Live Cells ◽  
Membrane Diffusion ◽  
Surface Receptor ◽  
Cellular Compartments ◽  
Sub Wavelength ◽  
The Way

Over the last decade, single-molecule detection (SMD) gave biologists a tool to turn their dream, to follow a single molecule in live cells, into reality. SMD provides the advantages of identifying subpopulations and of localizing molecules with sub-wavelength precision. The use of nanometre-sized ligand–fluorophore complexes has even made it possible to track targets within confined cellular compartments. In this review, we first describe the main benefits of SMD in cellular imaging. We then show how SMD was used to unravel the membrane diffusion of glutamatergic receptors and how it sheds light on the way neurons can regulate membrane distribution of receptors.

scholarly journals A controlled T7 transcription-driven symmetric amplification cascade machinery for single-molecule detection of multiple repair glycosylases

2021 ◽  
Author(s):  
Li-juan Wang ◽  
Le Liang ◽  
Bing-jie Liu ◽  
BingHua Jiang ◽  
Chun-yang Zhang
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection ◽  
Amplification Cascade

A controlled T7 transcription-driven symmetric amplification cascade machinery is developed for single-molecule detection of multiple repair glycosylases.

Highly Sensitive Detection of Paraquat with Pillar[5]arene as an aptamer in α-Hemolysin Nanopore

2021 ◽  
Author(s):  
Xiaojia Jiang ◽  
Mingsong Zang ◽  
Fei Li ◽  
Chunxi Hou ◽  
Quan Luo ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Single Molecule ◽  
Single Molecule Detection ◽  
Sensitive Detection ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
The Real ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

Biological nanopore-based techniques have attracted more and more attention recently in the field of single-molecule detection, because they allow the real-time, sensitive, high-throughput analysis. Herein, we report an engineered biological...

scholarly journals Nanopore Technology for the Application of Protein Detection

Nanomaterials ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1942
Author(s):  
Xiaoqing Zeng ◽  
Yang Xiang ◽  
Qianshan Liu ◽  
Liang Wang ◽  
Qianyun Ma ◽  
...  
Keyword(s):  
Single Molecule ◽  
Protein Detection ◽  
High Sensitivity ◽  
Single Molecule Detection ◽  
Sequence Detection ◽  
Fast Detection ◽  
Material Basis ◽  
Sensing Technology ◽  
Detection Technology ◽  
Structure Recognition

Protein is an important component of all the cells and tissues of the human body and is the material basis of life. Its content, sequence, and spatial structure have a great impact on proteomics and human biology. It can reflect the important information of normal or pathophysiological processes and promote the development of new diagnoses and treatment methods. However, the current techniques of proteomics for protein analysis are limited by chemical modifications, large sample sizes, or cumbersome operations. Solving this problem requires overcoming huge challenges. Nanopore single molecule detection technology overcomes this shortcoming. As a new sensing technology, it has the advantages of no labeling, high sensitivity, fast detection speed, real-time monitoring, and simple operation. It is widely used in gene sequencing, detection of peptides and proteins, markers and microorganisms, and other biomolecules and metal ions. Therefore, based on the advantages of novel nanopore single-molecule detection technology, its application to protein sequence detection and structure recognition has also been proposed and developed. In this paper, the application of nanopore single-molecule detection technology in protein detection in recent years is reviewed, and its development prospect is investigated.

scholarly journals Motional dynamics of single Patched1 molecules in cilia are controlled by Hedgehog and cholesterol

2019 ◽  
Vol 116 (12) ◽  
pp. 5550-5557 ◽  
Author(s):  
Lucien E. Weiss ◽  
Ljiljana Milenkovic ◽  
Joshua Yoon ◽  
Tim Stearns ◽  
W. E. Moerner
Keyword(s):  
Single Molecule ◽  
Hedgehog Signaling ◽  
Primary Cilia ◽  
Transmembrane Protein ◽  
Cellular Level ◽  
Live Cells ◽  
Cholesterol Depletion ◽  
Motional Dynamics ◽  
Directional Transport ◽  
Bulk Behavior

The Hedgehog-signaling pathway is an important target in cancer research and regenerative medicine; yet, on the cellular level, many steps are still poorly understood. Extensive studies of the bulk behavior of the key proteins in the pathway established that during signal transduction they dynamically localize in primary cilia, antenna-like solitary organelles present on most cells. The secreted Hedgehog ligand Sonic Hedgehog (SHH) binds to its receptor Patched1 (PTCH1) in primary cilia, causing its inactivation and delocalization from cilia. At the same time, the transmembrane protein Smoothened (SMO) is released of its inhibition by PTCH1 and accumulates in cilia. We used advanced, single molecule-based microscopy to investigate these processes in live cells. As previously observed for SMO, PTCH1 molecules in cilia predominantly move by diffusion and less frequently by directional transport, and spend a fraction of time confined. After treatment with SHH we observed two major changes in the motional dynamics of PTCH1 in cilia. First, PTCH1 molecules spend more time as confined, and less time freely diffusing. This result could be mimicked by a depletion of cholesterol from cells. Second, after treatment with SHH, but not after cholesterol depletion, the molecules that remain in the diffusive state showed a significant increase in the diffusion coefficient. Therefore, PTCH1 inactivation by SHH changes the diffusive motion of PTCH1, possibly by modifying the membrane microenvironment in which PTCH1 resides.

Single-Molecule Detection by Laser-Induced Fluorescence Technique with a Position-Sensitive Photon-Counting Apparatus

1994 ◽  
Vol 33 (Part 1, No. 3A) ◽  
pp. 1571-1576 ◽  
Author(s):  
Mitsuru Ishikawa ◽  
Ken-ichi Hirano ◽  
Tsuyoshi Hayakawa ◽  
Shigeru Hosoi ◽  
Sydney Brenner
Keyword(s):  
Single Molecule ◽  
Photon Counting ◽  
Laser Induced Fluorescence ◽  
Single Molecule Detection ◽  
Fluorescence Technique ◽  
Position Sensitive
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