In Situ Amplification‐Based Imaging of RNA in Living Cells

2019 ◽  
Vol 131 (34) ◽  
pp. 11698-11709 ◽  
Author(s):  
Zhihe Qing ◽  
Jingyuan Xu ◽  
Jinlei Hu ◽  
Jing Zheng ◽  
Lei He ◽  
...  
Keyword(s):  
Living Cells

DNA nanolantern-based split aptamer probes for in-situ ATP imaging in living cells and lighting up mitochondria

The Analyst ◽  
2021 ◽  
Author(s):  
Yaxin Wang ◽  
Dong-Xia Wang ◽  
Jia-Yi Ma ◽  
Jing Wang ◽  
Yichen Du ◽  
...  
Keyword(s):  
Adenosine Triphosphate ◽  
Cell Metabolism ◽  
Living Cells ◽  
Expression Level ◽  
Specific Analysis

Accurate and specific analysis of adenosine triphosphate (ATP) expression level in living cells can provide valuable information for understanding cell metabolism, physiological activities and pathologic mechanism. Herein, DNA nanolantern-based split...

scholarly journals Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes.

1993 ◽  
Vol 123 (2) ◽  
pp. 431-441 ◽  
Author(s):  
K Ainger ◽  
D Avossa ◽  
F Morgan ◽  
S J Hill ◽  
C Barry ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Living Cells ◽  
Branch Points ◽  
Rna Granules ◽  
Intracellular Movement ◽  
Directional Movement ◽  
Ionic Detergent ◽  
Mrna Granules

We have studied transport and localization of MBP mRNA in oligodendrocytes in culture by microinjecting labeled mRNA into living cells and analyzing the intracellular distribution of the injected RNA by confocal microscopy. Injected mRNA initially appears dispersed in the perikaryon. Within minutes, the RNA forms granules which, in the case of MBP mRNA, are transported down the processes to the periphery of the cell where the distribution again becomes dispersed. In situ hybridization shows that endogenous MBP mRNA in oligodendrocytes also appears as granules in the perikaryon and processes and dispersed in the peripheral membranes. The granules are not released by extraction with non-ionic detergent, indicating that they are associated with the cytoskeletal matrix. Three dimensional visualization indicates that MBP mRNA granules are often aligned in tracks along microtubules traversing the cytoplasm and processes. Several distinct patterns of granule movement are observed. Granules in the processes undergo sustained directional movement with a velocity of approximately 0.2 micron/s. Granules at branch points undergo oscillatory motion with a mean displacement of 0.1 micron/s. Granules in the periphery of the cell circulate randomly with a mean displacement of approximately 1 micron/s. The results are discussed in terms of a multi-step pathway for transport and localization of MBP mRNA in oligodendrocytes. This work represents the first characterization of intracellular movement of mRNA in living cells, and the first description of the role of RNA granules in transport and localization of mRNA in cells.

Intracellular Nonenzymatic In Situ Growth of Three-Dimensional DNA Nanostructures for Imaging Specific Biomolecules in Living Cells

ACS Nano ◽  
2020 ◽  
Vol 14 (8) ◽  
pp. 9572-9584
Author(s):  
Ran Liu ◽  
Songbai Zhang ◽  
Ting-Ting Zheng ◽  
Yan-Ru Chen ◽  
Jing-Ting Wu ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Living Cells ◽  
In Situ Growth ◽  
Dna Nanostructures

Detecting Early Apoptosis in Whole Epithelial Sheets with a Caspase 3 Substrate, Phiphilux and Annexes V Using Confocal Microscopy

2001 ◽  
Vol 7 (S2) ◽  
pp. 600-601
Author(s):  
Kathy K. H. Svoboda
Keyword(s):  
In Situ Hybridization ◽  
Confocal Microscopy ◽  
General Principle ◽  
Caspase 3 ◽  
Living Cells ◽  
Corneal Epithelial ◽  
Early Apoptosis ◽  
Epithelial Sheets ◽  
Conventional Light Microscope

Many reagents have been developed recently to label living cells with substrates that will become fluorescent if an enzyme is active. The general principle is that the substrate will be taken up by living cells then detected only if the enzyme is active. These substrates work well with isolated individual cells, however, more difficulty can be encountered when studying whole tissues. Problems can range from substrate penetration into whole tissues to being able to detect the label effectively. We have used the chicken corneal epithelia for many studies, but the tissue is to thick to view with a conventional light microscope, therefore we have developed techniques using laser confocal microscopes to view this tissue in with a variety of techniques including in situ hybridization, immunohistochemistry, and vital dyes/stains.Whole embryonic corneal epithelial sheets can be isolated without the basal lamina.

scholarly journals Studies using fluorescent tetrahydrochrysene estrogens for in situ visualization of the estrogen receptor in living cells.

1995 ◽  
Vol 9 (5) ◽  
pp. 592-604
Author(s):  
R J Miksicek ◽  
K E Carlson ◽  
K J Hwang ◽  
J A Katzenellenbogen
Keyword(s):  
Estrogen Receptor ◽  
Living Cells ◽  
In Situ Visualization

Experiments in Nanomechanical Properties of Live Osteoblast Cells and Cell–Biomaterial Interface

2011 ◽  
Vol 2 (4) ◽  
Author(s):  
Rohit Khanna ◽  
Kalpana S. Katti ◽  
Dinesh R. Katti
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Cell Structure ◽  
Living Cells ◽  
Actin Stress Fiber ◽  
Cell Culture Medium ◽  
Nanomechanical Properties ◽  
Cell Substrate ◽  
Significant Difference

Characterizing the mechanical characteristics of living cells and cell–biomaterial composite is an important area of research in bone tissue engineering. In this work, an in situ displacement-controlled nanoindentation technique (using Hysitron Triboscope) is developed to perform nanomechanical characterization of living cells (human osteoblasts) and cell–substrate constructs under physiological conditions (cell culture medium; 37 °C). In situ elastic moduli (E) of adsorbed proteins on tissue culture polystyrene (TCPS) under cell culture media were found to be ∼4 GPa as revealed by modulus mapping experiments. The TCPS substrates soaked in cell culture medium showed significant difference in surface nanomechanical properties (up to depths of ∼12 nm) as compared to properties obtained from deeper indentations. Atomic force microscopy (AFM) revealed the cytoskeleton structures such as actin stress fiber networks on flat cells which are believed to impart the structural integrity to cell structure. Load-deformation response of cell was found to be purely elastic in nature, i.e., cell recovers its shape on unloading as indicated by linear loading and unloading curves obtained at 1000 nm indentation depth. The elastic response of cells is obtained during initial cell adhesion (ECell, 1 h, 1000 nm = 4.4–12.4 MPa), cell division (ECell, 2 days, 1000 nm = 1.3–3.0 MPa), and cell spreading (ECell, 2 days, 1000 nm = 6.9–11.6 MPa). Composite nanomechanical responses of cell–TCPS constructs were obtained by indentation at depths of 2000 nm and 3000 nm on cell-seeded TCPS. Elastic properties of cell–substrate composites were mostly dominated by stiff TCPS (EBulk = 5 GPa) lying underneath the cell.

scholarly journals Electron Microscopy of Living Cells During in Situ Fluorescence Microscopy

ACS Nano ◽  
2015 ◽  
Vol 10 (1) ◽  
pp. 265-273 ◽  
Author(s):  
Nalan Liv ◽  
Daan S. B. van Oosten Slingeland ◽  
Jean-Pierre Baudoin ◽  
Pieter Kruit ◽  
David W. Piston ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Fluorescence Microscopy ◽  
Living Cells
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