Genetically Engineered Pores as Metal Ion Biosensors

1993 ◽  
Vol 330 ◽  
Author(s):  
John Kasianowicz ◽  
Barbara Walker ◽  
Musti Krishnasastry ◽  
Hagan Bayley

ABSTRACTWe are adapting proteins that form pores in lipid bilayers for use as components of biosensors. Specifically, we have produced genetically engineered variants of the α hemolysin (αHL) fromStaphylococcusaureus with properties that are sensitive to low concentrations of divalent cations. For example, the pore-forming activity of one mutant (αHL-H5: residues 130–134 inclusive replaced with histidine) is inhibited by Zn2+at concentrations as low as 1 μM, as judged by the reduction in its ability to lyse rabbit red blood cells and to increase the conductance of planar lipid bilayer membranes. When αHL-H5 is added to the aqueous phase bathing one side of a planar membrane, the subsequent addition of 100 μM Zn2+to either side blocks the pores that form. This result suggests that at least part of the mutated region lines the channel lumen. Ca2+and Mg2+do not block the channel and therefore the H5 mutation confers a degree of analyte specificity to the αHL pore. The results suggest that genetically engineered pores have great promise for the rapid and sensitive detection of metal cations and we discuss the merits and potential limitations for their use in this application. Specifically, we examine the issues of selectivity, sensitivity, response time, dynamic range and longevity. Some of these properties are interdependent. For example, the goals of high sensitivity and rapid response time can be in conflict.

2021 ◽  
Author(s):  
Milena Timcenko ◽  
Anton A.A. Autzen ◽  
Henriette E. Autzen

Amphiphilic copolymers show great promise in extracting membrane proteins directly from lipid bilayers into 'native nanodiscs'. However, many such copolymers are polyanionic and sensitive to divalent cations, limiting their applicability towards Ca2+ or Mg2+ dependent proteins. Here, we characterize the Ca2+ and Mg2+ sensitivity of poly(acrylic acid-co-styrene) (AASTY) copolymers using analytical UV and fluorescent size exclusion chromatography, enabling us to separate signals from nanodiscs, copolymers, and soluble aggregates. Determination of free Ca2+ ion concentrations in the presence of copolymer shows that divalent cation tolerance is dependent on not only specific characteristics of a copolymer, but also on its concentration. We see that high ionic strength protects against aggregation facilitated by divalent cations, which is prominent in nanodiscs isolated from excess free copolymer through dialysis. Overall, we conclude that the behavior of amphiphilic copolymers in the presence of divalent cations is more complex than precipitation beyond a specific cation concentration.


2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Omer Isildak ◽  
Furkan Saymaz ◽  
Ahmet Karadag ◽  
Nesrin Okumus Korkmaz ◽  
Azade Attar

A novel potentiometric sensor based on ionophore (Cd(NH2CH2CH2OCH2CH2OCH2CH2NH2)Ag3(CN)5) for the determination ofβ-N-oxalyl-l-α,β-diaminopropionic acid (ODAP) is developed. The ODAP-selective membrane sensor demonstrates high sensitivity and short response time. The detection limit of the ODAP-selective membrane sensor is about2×10-6 mol L-1and the response time is shorter than 6 s. The linear dynamic range of the ODAP-selective membrane sensor is between ODAP concentrations of1.0×10-2and1×10-6 mol L-1. The ODAP-selective membrane sensor exhibits good operational stability for at least one week in dry conditions at 4–6°C. It has a reproducible and stable response during continuous work for at least 10 h with a relative standard deviation of 0.28% (n= 18).


Author(s):  
F. Ouyang ◽  
D. A. Ray ◽  
O. L. Krivanek

Electron backscattering Kikuchi diffraction patterns (BKDP) reveal useful information about the structure and orientation of crystals under study. With the well focused electron beam in a scanning electron microscope (SEM), one can use BKDP as a microanalysis tool. BKDPs have been recorded in SEMs using a phosphor screen coupled to an intensified TV camera through a lens system, and by photographic negatives. With the development of fiber-optically coupled slow scan CCD (SSC) cameras for electron beam imaging, one can take advantage of their high sensitivity and wide dynamic range for observing BKDP in SEM.We have used the Gatan 690 SSC camera to observe backscattering patterns in a JEOL JSM-840A SEM. The CCD sensor has an active area of 13.25 mm × 8.83 mm and 576 × 384 pixels. The camera head, which consists of a single crystal YAG scintillator fiber optically coupled to the CCD chip, is located inside the SEM specimen chamber. The whole camera head is cooled to about -30°C by a Peltier cooler, which permits long integration times (up to 100 seconds).


Author(s):  
N. Mori ◽  
T. Oikawa ◽  
Y. Harada ◽  
J. Miyahara ◽  
T. Matsuo

The Imaging Plate (IP) is a new type imaging device, which was developed for diagnostic x ray imaging. We have reported that usage of the IP for a TEM has many merits; those are high sensitivity, wide dynamic range, and good linearity. However in the previous report the reading system was prototype drum-type-scanner, and IP was also experimentally made, which phosphor layer was 50μm thick with no protective layer. So special care was needed to handle them, and they were used only to make sure the basic characteristics. In this article we report the result of newly developed reading, printing system and high resolution IP for practical use. We mainly discuss the characteristics of the IP here. (Precise performance concerned with the reader and other system are reported in the other article.)Fig.1 shows the schematic cross section of the IP. The IP consists of three parts; protective layer, phosphor layer and support.


2020 ◽  
Vol 39 (1) ◽  
pp. 231-246 ◽  
Author(s):  
Xian Zheng ◽  
Wenyu Cheng ◽  
Chendong Ji ◽  
Jin Zhang ◽  
Meizhen Yin

Abstract Metal ions are widely present in biological systems and participate in many critical biochemical processes such as material transportation, energy conversion, information transmission and metabolic regulation, making them indispensable substance in our body. They can cause health problems when deficiency or excess occurs. To understand various metabolic processes and facilitate diseases diagnosis, it is very important to measure the content and monitor the distribution of metal ions in individual cells, tissues and whole organisms. Among the various methods for metal ion detection, fluorescent sensors with organic dyes have attracted tremendous attention due to many advantages such as high fluorescence quantum yield, facile modification approaches and biocompatibility in addition to operation ease, high sensitivity, fast detection speed, and real-time detection. This review summarizes the recent progress on the detection and imaging of the metal ions in biological systems including Na+, K+, Ca2+, Mg2+, Fe2+/Fe3+, Zn2+, and Cu2+ provides an opinion on remaining challenges to be addressed in this field.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Libo Zhang ◽  
Zhiqingzi Chen ◽  
Kaixuan Zhang ◽  
Lin Wang ◽  
Huang Xu ◽  
...  

AbstractThe advent of topological semimetals enables the exploitation of symmetry-protected topological phenomena and quantized transport. Here, we present homogeneous rectifiers, converting high-frequency electromagnetic energy into direct current, based on low-energy Dirac fermions of topological semimetal-NiTe2, with state-of-the-art efficiency already in the first implementation. Explicitly, these devices display room-temperature photosensitivity as high as 251 mA W−1 at 0.3 THz in an unbiased mode, with a photocurrent anisotropy ratio of 22, originating from the interplay between the spin-polarized surface and bulk states. Device performances in terms of broadband operation, high dynamic range, as well as their high sensitivity, validate the immense potential and unique advantages associated to the control of nonequilibrium gapless topological states via built-in electric field, electromagnetic polarization and symmetry breaking in topological semimetals. These findings pave the way for the exploitation of topological phase of matter for high-frequency operations in polarization-sensitive sensing, communications and imaging.


Coatings ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 685
Author(s):  
Ai-Huei Chiou ◽  
Jun-Luo Wei ◽  
Ssu-Han Chen

A novel surface-enhanced Raman scattering (SERS)-based probe to capture heavy metal ion (Zn2+) by bovine serum albumin (BSA) using Si-nanowire (SiNW) arrays with silver nanoparticles (AgNPs) was developed. A layer with AgNPs was deposited on the SiNW surface by RF magnetron sputtering for enhancement of SERS signals. Using a high-resolution transmission electron microscope (HRTEM), the observation reveals that the AgNP layer with depths of 30–75 nm was successfully deposited on SiNW arrays. The Ag peaks in EDS and XRD spectra of SiNW arrays confirmed the presence of Ag particles on SiNW arrays. The WCA observations showed a high affinity of the Ag–SiNW arrays immobilized with BSA (water contact angle (WCA) = 87.1°) and ZnSO4 (WCA = 8.8°). The results of FTIR analysis illustrate that the conjugate bonds exist between zinc sulfate (ZnSO4) and –OH groups/–NH groups of BSA. The resulting SiNWs/Ag NPs composite interfaces showed large Raman scattering enhancement for the capture of heavy metal ions by BSA with a detection of 0.1 μM. BSA and ZnSO4 conjugations, illustrating specific SERS spectra with high sensitivity, which suggests great promise in developing label-free biosensors.


2002 ◽  
Vol 38 (13) ◽  
pp. 650 ◽  
Author(s):  
H. Matsuda ◽  
A. Miura ◽  
H. Irie ◽  
S. Tanaka ◽  
K. Ito ◽  
...  

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