scholarly journals Use of Novel Mutant Galactosyltransferase for the Bioconjugation of Terminal N-Acetylglucosamine (GlcNAc) Residues on Live Cell Surface

2013 ◽  
Vol 24 (1) ◽  
pp. 144-152 ◽  
Author(s):  
Natalia Mercer ◽  
Boopathy Ramakrishnan ◽  
Elizabeth Boeggeman ◽  
Luke Verdi ◽  
Pradman K. Qasba
Keyword(s):  
Cell Surface ◽  
Live Cell

scholarly journals Cell Surface Receptors: Rapid Quantification of Live Cell Receptors Using Bioluminescence in a Flow-Based Microfluidic Device (Small 8/2015)

Small ◽  
2015 ◽  
Vol 11 (8) ◽  
pp. 1012-1012
Author(s):  
Ramesh Ramji ◽  
Cheong Fook Cheong ◽  
Hiroaki Hirata ◽  
Abdur Rub Abdur Rahman ◽  
Chwee Teck Lim
Keyword(s):  
Cell Surface ◽  
Microfluidic Device ◽  
Live Cell ◽  
Cell Surface Receptors ◽  
Cell Receptors ◽  
Surface Receptors ◽  
Rapid Quantification

Generation of Live-Cell Microarrays by Means of DNA-Directed Immobilization of Specific Cell-Surface Ligands

2007 ◽  
Vol 46 (22) ◽  
pp. 4180-4183 ◽  
Author(s):  
Hendrik Schroeder ◽  
Bernhard Ellinger ◽  
Christian F. W. Becker ◽  
Herbert Waldmann ◽  
Christof M. Niemeyer
Keyword(s):  
Cell Surface ◽  
Live Cell ◽  
Specific Cell ◽  
Surface Ligands ◽  
Cell Microarrays ◽  
Specific Cell Surface

scholarly journals Golgi, trafficking, and mitosis dysfunctions in pulmonary arterial endothelial cells exposed to monocrotaline pyrrole and NO scavenging

2009 ◽  
Vol 297 (4) ◽  
pp. L715-L728 ◽  
Author(s):  
Jason Lee ◽  
Reuben Reich ◽  
Fang Xu ◽  
Pravin B. Sehgal
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Cell Surface ◽  
Live Cell ◽  
Cell Uptake ◽  
Important Distinction ◽  
No Donor ◽  
Pulmonary Arterial ◽  
Monocrotaline Pyrrole ◽  
Arterial Endothelial Cells

Although the administration of monocrotaline (MCT) into experimental animals is in widespread use today in investigations of pulmonary arterial hypertension (PAH), the underlying cellular and subcellular mechanisms that culminate in vascular remodeling are incompletely understood. Bovine pulmonary arterial endothelial cells (PAECs) in culture exposed to monocrotaline pyrrole (MCTP) develop “megalocytosis” 18–24 h later characterized by enlarged hyperploid cells with enlarged Golgi, mislocalization of endothelial nitric oxide synthase away from the plasma membrane, decreased cell-surface/caveolar nitric oxide (NO), and hypo- S-nitrosylation of caveolin-1, clathrin heavy chain, and N-ethylmaleimide-sensitive factor. We investigated whether MCTP did in fact affect functional intracellular trafficking. The NO scavenger (4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO) and the NO donor diethylamine NONOate were used for comparison. Both MCTP and c-PTIO produced distinctive four- to fivefold enlarged PAECs within 24–48 h with markedly enlarged/dispersed Golgi, as visualized by immunostaining for the Golgi tethers/matrix proteins giantin, GM130, and p115. Live-cell uptake of the Golgi marker C5 ceramide revealed a compact juxtanuclear Golgi in untreated PAECs, brightly labeled enlarged circumnuclear Golgi after MCTP, but minimally labeled Golgi elements after c-PTIO. These Golgi changes were reduced by NONOate. After an initial inhibition during the first day, both MCTP and c-PTIO markedly enhanced anterograde secretion of soluble cargo (exogenous vector-expressed recombinant horseradish peroxidase) over the next 4 days. Live-cell internalization assays using fluorescently tagged ligands showed that both MCTP and c-PTIO inhibited the retrograde uptake of acetylated low-density lipoprotein, transferrin, and cholera toxin B. Moreover, MCTP, and to a variable extent c-PTIO, reduced the cell-surface density of all receptors assayed (LDLR, TfnR, BMPR, Tie-2, and PECAM-1/CD31). In an important distinction, c-PTIO enhanced mitosis in PAECs but MCTP inhibited mitosis, even that due to c-PTIO, despite markedly exaggerated Golgi dispersal. Taken together, these data define a broad-spectrum Golgi and subcellular trafficking dysfunction syndrome in endothelial cells exposed to MCTP or NO scavenging.

scholarly journals Live Cell Surface Conjugation Methods for Imaging, Sensing and Therapy

2018 ◽  
Vol 114 (3) ◽  
pp. 20a ◽  
Author(s):  
Joydeb Majumder ◽  
Gaurav Chopra
Keyword(s):  
Cell Surface ◽  
Live Cell

scholarly journals Nano-Scale Topography Imaging of Live Cell Surface Using Scanning Ion Conductance Microscopy

Hyomen Kagaku ◽  
2015 ◽  
Vol 36 (6) ◽  
pp. 313-318 ◽  
Author(s):  
Hiroki IDA ◽  
Yasufumi TAKAHASHI ◽  
Hitoshi SHIKU ◽  
Tomokazu MATSUE
Keyword(s):  
Cell Surface ◽  
Live Cell ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Nano Scale

scholarly journals Immunotargeting of Nanocrystals by SpyCatcher Conjugation of Engineered Antibodies

2021 ◽  
Author(s):  
Cassio Pedroso ◽  
Victor Mann ◽  
Kathrin Zuberbühler ◽  
Markus-Frederik Bohn ◽  
Jessica Yu ◽  
...  
Keyword(s):  
Cell Surface ◽  
Specific Binding ◽  
Time Lapse ◽  
Live Cell ◽  
Confocal Imaging ◽  
Cellular Targets ◽  
Type Plasminogen Activator ◽  
Urokinase Type Plasminogen Activator ◽  
Isopeptide Bonds ◽  
Time Lapse Imaging

Inorganic nanocrystals such as quantum dots (QDs) and upconverting nanoparticles (UCNPs) are uniquely suited for quanti-tative live-cell imaging and are typically functionalized with ligands to study specific receptors or cellular targets. Antibod-ies (Ab) are among the most useful targeting reagents owing to their high affinities and specificities, but common nanocrys-tal labeling methods may orient Ab incorrectly, be reversible or denaturing, or lead to Ab-NP complexes too large for some applications. Here, we show that SpyCatcher proteins, which bind and spontaneously form covalent isopeptide bonds with cognate SpyTag peptides, can conjugate engineered Ab to nanoparticle surfaces with control over stability, orientation, and stoichiometry. Compact SpyCatcher-functionalized QDs and UCNPs may be labeled with short-chain variable fragment Ab (scFv) engineered to bind urokinase-type plasminogen activator receptors (uPAR) that are overexpressed in many human can-cers. Confocal imaging of anti-uPAR scFv-QD conjugates shows the Ab mediates specific binding and internalization by breast cancer cells expressing uPAR. Time-lapse imaging of photostable scFv-UCNP conjugates show that Ab binding caus-es uPAR internalization with a ∼20-minute half-life on the cell surface, and uPAR is internalized to endolysosomal com-partments distinct from general membrane stains and without significant recycling to the cell surface. The controlled and stable conjugation of engineered Ab to NPs enables targeting of diverse receptors for live-cell study of their distribution, trafficking, and physiology.

Rolling “wool-balls”: rapid live-cell mapping of membrane sialic acids via poly-p-benzoquinone/ethylenediamine nanoclusters

2019 ◽  
Vol 55 (65) ◽  
pp. 9681-9684 ◽  
Author(s):  
Bin-Bin Chen ◽  
Xiao-Yuan Wang ◽  
Ruo-Can Qian
Keyword(s):  
Cell Surface ◽  
Sialic Acids ◽  
Live Cell ◽  
Cell Mapping

In this work, we design sticky, furry and fluorescent “wool-balls” based on p-benzoquinone/ethylenediamine polymer nanoclusters, which provide a convenient, fast labeling strategy for the imaging of cell surface sialic acids.

Flow cytometry and live cell imaging for characterization of cell surface markers on human tissue-derived progenitors

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S69-S70
Author(s):  
W.A. Bova ◽  
V.R. Mantripragada ◽  
V. Luangphakdy ◽  
G.F. Muschler
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Human Tissue ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Surface Markers ◽  
Cell Surface Markers
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