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 Adenosine Triphosphate Measurement in Deep Sea Using a Microfluidic Device

Micromachines ◽  
2018 ◽  
Vol 9 (8) ◽  
pp. 370 ◽  
Author(s):  
Tatsuhiro Fukuba ◽  
Takuroh Noguchi ◽  
Kei Okamura ◽  
Teruo Fujii
Keyword(s):  
Adenosine Triphosphate ◽  
Microfluidic Device ◽  
Deep Sea ◽  
Temperature Sensor ◽  
Biochemical Parameter ◽  
Underwater Vehicle ◽  
Luciferase Assay ◽  
Bioluminescence Intensity

Total ATP (adenosine triphosphate) concentration is a useful biochemical parameter for detecting microbial biomass or biogeochemical activity anomalies in the natural environment. In this study, we describe the development and evaluation of a new version of in situ ATP analyzer improved for the continuous and quantitative determination of ATP in submarine environments. We integrated a transparent microfluidic device containing a microchannel for cell lysis and a channel for the bioluminescence L–L (luciferin–luciferase) assay with a miniature pumping unit and a photometry module for the measurement of the bioluminescence intensity. A heater and a temperature sensor were also included in the system to maintain an optimal temperature for the L–L reaction. In this study, the analyzer was evaluated in deep sea environments, reaching a depth of 200 m using a remotely operated underwater vehicle. We show that the ATP analyzer successfully operated in the deep-sea environment and accurately quantified total ATP within the concentration lower than 5 × 10−11 M.

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.

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

Cell Metabolism of the Overloaded Mammalian Heart in Situ

Cardiology ◽  
1959 ◽  
Vol 34 (1) ◽  
pp. 19-27 ◽  
Author(s):  
L. Szekeres ◽  
M. Schein
Keyword(s):  
Cell Metabolism ◽  
Mammalian Heart

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.

NMR methods for in-situ biofilm metabolism studies: spatial and temporal resolved measurements

2005 ◽  
Vol 52 (7) ◽  
pp. 7-12 ◽  
Author(s):  
P.D. Majors ◽  
J.S. McLean ◽  
J.K. Fredrickson ◽  
R.A. Wind
Keyword(s):  
Magnetic Resonance ◽  
Cell Metabolism ◽  
Shewanella Oneidensis ◽  
Growth Conditions ◽  
Magnetic Resonance Images ◽  
Ex Situ ◽  
Adherent Cell ◽  
Growth Environment ◽  
Environment Control

We are developing novel nuclear magnetic resonance (NMR) microscopy, spectroscopy and combined NMR/optical techniques for the study of biofilms under known, controlled growth conditions. Objectives include: time and depth-resolved metabolite concentrations with isotropic spatial resolution on the order of 10 microns, metabolic pathways and flux rates, mass transport and ultimately their correlation with gene expression by optical microscopy in biofilms. We describe the implementation of ex-situ grown biofilms to improve growth environment control and NMR analysis. In-situ NMR depth resolved metabolite profiling techniques are introduced and demonstrated for a Shewanella oneidensis strain MR-1 biofilm. Finally, initial combined confocal fluorescence and magnetic resonance images are shown for a GFP-labeled Shewanella biofilm. These methods are equally applicable to other biofilm systems of interest; thus they may provide a significant contribution toward the understanding of adherent cell metabolism.

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