Asymmetric Nanopore Electrode-Based Amplification for Electron Transfer Imaging in Live Cells

2018 ◽  
Vol 140 (16) ◽  
pp. 5385-5392 ◽  
Author(s):  
Yi-Lun Ying ◽  
Yong-Xu Hu ◽  
Rui Gao ◽  
Ru-Jia Yu ◽  
Zhen Gu ◽  
...  
2019 ◽  
Vol 116 (3) ◽  
pp. 315a
Author(s):  
Yilun Ying ◽  
Yongxu Hu ◽  
Rui Gao ◽  
Ling-Fei Cui ◽  
Yi-Tao Long

2016 ◽  
Vol 40 (7) ◽  
pp. 6101-6108 ◽  
Author(s):  
Turibius Simon ◽  
Muthaiah Shellaiah ◽  
Venkatesan Srinivasadesikan ◽  
Ching-Chang Lin ◽  
Fu-Hsiang Ko ◽  
...  

A simple anthracene-based AP probe was synthesized to detect Cu2+ ions, via the photoinduced electron transfer mechanism, in live cells.


2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Hongbao Xin ◽  
Wen Jing Sim ◽  
Bumseok Namgung ◽  
Yeonho Choi ◽  
Baojun Li ◽  
...  

2014 ◽  
Vol 86 (9) ◽  
pp. 4395-4401 ◽  
Author(s):  
Jie Zhou ◽  
Chuanan Liao ◽  
Limin Zhang ◽  
Qigang Wang ◽  
Yang Tian
Keyword(s):  

2021 ◽  
Author(s):  
Caleb Hoopes ◽  
Francisco Garcia ◽  
Akash Sarkar ◽  
Nicholas Kuehl ◽  
David Barkan ◽  
...  

Tryptophan (Trp) plays a variety of critical functional roles in protein biochemistry however, owing to its low natural frequency and poor nucleophilicity, the design of effective methods for both single protein bioconjugation at Trp as well as for in situ chemoproteomic profiling re-mains a challenge. Here, we report a method for covalent Trp modification that is suitable for both scenarios by invoking photo-induced electron transfer (PET) as a means of driving efficient reactivity. We have engineered biaryl N-carbamoyl pyridinium salts that possess a donor-acceptor relationship enabling optical triggering with visible light whilst simultaneously attenuating the probe’s photo-oxidation potential in order to prevent photodegradation. This probe was assayed against a small bank of eight peptides and proteins, where it was found that micromolar concentrations of probe and short irradiation times (10-60 min) with violet light enabled efficient reactivity towards surface exposed Trp residues. The carbamate transferring group can be used to transfer useful functional groups to proteins including affinity tags and click handles. DFT calculations and other mechanistic analyses reveal correlations between excited state lifetimes, relative fluorescent quantum yields, and chemical reactivity. Biotinylated and azide-functionalized pyridinium salts were used for Trp profiling in HEK293T lysates and in situ in HEK293T cells using 450 nm LED irradiation. Peptide level enrichment from live cell labelling experiments identified 290 Trp modifications, with an 82% selectivity for Trp modification over other π-amino acids; demonstrating the ability of this method to identify and quantify reactive Trp residues from live cells.


2007 ◽  
Vol 74 (3) ◽  
pp. 615-623 ◽  
Author(s):  
Harald von Canstein ◽  
Jun Ogawa ◽  
Sakayu Shimizu ◽  
Jonathan R. Lloyd

ABSTRACT Fe(III)-respiring bacteria such as Shewanella species play an important role in the global cycle of iron, manganese, and trace metals and are useful for many biotechnological applications, including microbial fuel cells and the bioremediation of waters and sediments contaminated with organics, metals, and radionuclides. Several alternative electron transfer pathways have been postulated for the reduction of insoluble extracellular subsurface minerals, such as Fe(III) oxides, by Shewanella species. One such potential mechanism involves the secretion of an electron shuttle. Here we identify for the first time flavin mononucleotide (FMN) and riboflavin as the extracellular electron shuttles produced by a range of Shewanella species. FMN secretion was strongly correlated with growth and exceeded riboflavin secretion, which was not exclusively growth associated but was maximal in the stationary phase of batch cultures. Flavin adenine dinucleotide was the predominant intracellular flavin but was not released by live cells. The flavin yields were similar under both aerobic and anaerobic conditions, with total flavin concentrations of 2.9 and 2.1 μmol per gram of cellular protein, respectively, after 24 h and were similar under dissimilatory Fe(III)-reducing conditions and when fumarate was supplied as the sole electron acceptor. The flavins were shown to act as electron shuttles and to promote anoxic growth coupled to the accelerated reduction of poorly crystalline Fe(III) oxides. The implications of flavin secretion by Shewanella cells living at redox boundaries, where these mineral phases can be significant electron acceptors for growth, are discussed.


Author(s):  
P. Bonhomme ◽  
A. Beorchia

We have already described (1.2.3) a device using a pockel's effect light valve as a microscopical electron image converter. This converter can be read out with incoherent or coherent light. In the last case we can set in line with the converter an optical diffractometer. Now, electron microscopy developments have pointed out different advantages of diffractometry. Indeed diffractogram of an image of a thin amorphous part of a specimen gives information about electron transfer function and a single look at a diffractogram informs on focus, drift, residual astigmatism, and after standardizing, on periods resolved (4.5.6). These informations are obvious from diffractogram but are usualy obtained from a micrograph, so that a correction of electron microscope parameters cannot be realized before recording the micrograph. Diffractometer allows also processing of images by setting spatial filters in diffractogram plane (7) or by reconstruction of Fraunhofer image (8). Using Electrotitus read out with coherent light and fitted to a diffractometer; all these possibilities may be realized in pseudoreal time, so that working parameters may be optimally adjusted before recording a micrograph or before processing an image.


Author(s):  
Shinya Inoué

This paper reports progress of our effort to rapidly capture, and display in time-lapsed mode, the 3-dimensional dynamic architecture of active living cells and developing embryos at the highest resolution of the light microscope. Our approach entails: (A) real-time video tape recording of through-focal, ultrathin optical sections of live cells at the highest resolution of the light microscope; (B) repeat of A at time-lapsed intervals; (C) once each time-lapsed interval, an image at home focus is recorded onto Optical Disk Memory Recorder (OMDR); (D) periods of interest are selected using the OMDR and video tape records; (E) selected stacks of optical sections are converted into plane projections representing different view angles (±4 degrees for stereo view, additional angles when revolving stereos are desired); (F) analysis using A - D.


Author(s):  
E. D. Salmon ◽  
J. C. Waters ◽  
C. Waterman-Storer

We have developed a multi-mode digital imaging system which acquires images with a cooled CCD camera (Figure 1). A multiple band pass dichromatic mirror and robotically controlled filter wheels provide wavelength selection for epi-fluorescence. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. Many of our experiments involve investigations of spindle assembly dynamics and chromosome movements in live cells or unfixed reconstituted preparations in vitro in which photodamage and phototoxicity are major concerns. As a consequence, a major factor in the design was optical efficiency: achieving the highest image quality with the least number of illumination photons. This principle applies to both epi-fluorescence and transmitted light imaging modes. In living cells and extracts, microtubules are visualized using X-rhodamine labeled tubulin. Photoactivation of C2CF-fluorescein labeled tubulin is used to locally mark microtubules in studies of microtubule dynamics and translocation. Chromosomes are labeled with DAPI or Hoechst DNA intercalating dyes.


Sign in / Sign up

Export Citation Format

Share Document