In Situ Assay of Proteins Incorporated with Unnatural Amino Acids in Single Living Cells by Differenced Resonance Light Scattering Correlation Spectroscopy

2021 ◽  
Author(s):  
Jinchun Xu ◽  
Yaoqi Liu ◽  
Fucai Li ◽  
Liyun Deng ◽  
Chaoqing Dong ◽  
...  
Keyword(s):  
Amino Acids ◽  
Light Scattering ◽  
Unnatural Amino Acids ◽  
Living Cells ◽  
Resonance Light Scattering ◽  
Correlation Spectroscopy ◽  
Single Living Cells ◽  
Single Living ◽  
In Situ Assay

In situ investigations of single living cells infected by viruses

1992 ◽  
Vol 42-44 ◽  
pp. 1161-1167 ◽  
Author(s):  
W. Häberle ◽  
J.K.H. Hörber ◽  
F. Ohnesorge ◽  
D.P.E. Smith ◽  
G. Binnig
Keyword(s):  
Living Cells ◽  
Single Living Cells ◽  
Single Living

scholarly journals Insulin-triggered repositioning of munc18c on syntaxin-4 in GLUT4 signalling

2008 ◽  
Vol 410 (2) ◽  
pp. 255-260 ◽  
Author(s):  
Natalie P. Smithers ◽  
Conrad P. Hodgkinson ◽  
Matt Cuttle ◽  
Graham J. Sale
Keyword(s):  
Plasma Membrane ◽  
Glucose Transporter ◽  
Cell Signalling ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Vesicle Docking ◽  
Fluorescence Correlation ◽  
Syntaxin 4 ◽  
Single Living Cells ◽  
Single Living

One of the most important actions of insulin is the stimulation of the uptake of glucose into fat and muscle cells. Crucial to this response is the translocation of GLUT4 (glucose transporter-4) to the plasma membrane. The insulin-stimulated GLUT4 vesicle docking at the plasma membrane requires an interaction between VAMP-2 (vesicle-associated membrane protein-2) on the GLUT4 vesicle and syntaxin-4 in the plasma membrane. In the basal state, munc18c is thought to preclude GLUT4 vesicle docking by inhibiting this interaction. Here, we have used FCS (fluorescence correlation spectroscopy) in single living cells to show that munc18c binds to syntaxin-4 in both the basal and insulin-stimulated states. We show that munc18c contains two binding sites for syntaxin-4, one of which is disrupted by insulin, while the other is activated by insulin. Insulin-triggered repositioning of munc18c on syntaxin-4 in this way in turn allows syntaxin-4 to adopt its ‘open’ conformation and bind VAMP-2, resulting in the docking of the GLUT4 vesicle at the cell surface. The results also demonstrate the utility of using FCS in intact single living cells to elucidate cell signalling events.

Simultaneously monitoring endogenous MAPK members in single living cells by multi-channel fluorescence correlation spectroscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Liyun Deng ◽  
Xiangyi Huang ◽  
Chaoqing Dong ◽  
Jicun Ren
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Mapk Pathway ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Mitogen Activated Protein Kinase ◽  
Fluorescence Correlation ◽  
Mitogen Activated Protein ◽  
Cellular Signal ◽  
Single Living Cells ◽  
Single Living

The mitogen-activated protein kinase (MAPK) pathway is a major module for cellular signal transduction. The dysregulation of MAPK pathway has been involved in the pathogenesis of multiple diseases ranging from...

Application of FRAP and digitized fluorescence microscopy to problems in cell membrane dynamics

1988 ◽  
Vol 46 ◽  
pp. 118-119
Author(s):  
K. Jacobson ◽  
A. Ishihara ◽  
B. Holifield ◽  
F. Zhang
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Extended Period ◽  
Dynamic Structure ◽  
Living Cells ◽  
Membrane Dynamics ◽  
Molecular Cell Biology ◽  
Image Averaging ◽  
Single Living Cells ◽  
Single Living

Our laboratory is concerned with understanding the dynamic structure of the plasma membrane with particular reference to the movement of membrane constituents during cell locomotion. In addition to the standard tools of molecular cell biology, we employ both fluorescence recovery after photo- bleaching (FRAP) and digitized fluorescence microscopy (DFM) to investigate individual cells. FRAP allows the measurement of translational mobility of membrane and cytoplasmic molecules in small regions of single, living cells. DFM is really a new form of light microscopy in that the distribution of individual classes of ions, molecules, and macromolecules can be followed in single, living cells. By employing fluorescent antibodies to defined antigens or fluorescent analogs of cellular constituents as well as ultrasensitive, electronic image detectors and video image averaging to improve signal to noise, fluorescent images of living cells can be acquired over an extended period without significant fading and loss of cell viability.

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