A simple, sensitive and selective quantum-dot-based western blot method for the simultaneous detection of multiple targets from cell lysates

Quantum dots (Qdots) are fluorescent nanoparticles that have great potential as detection agents in biological applications. Their optical properties, including photostability and narrow, symmetrical emission bands with large Stokes shifts, and the potential for multiplexing of many different colours, give them significant advantages over traditionally used fluorescent dyes. Here, we report the straightforward generation of stable, covalent quantum dot-protein A/G bioconjugates that will be able to bind to almost any IgG antibody, and therefore can be used in many applications. An additional advantage is that the requirement for a secondary antibody is removed, simplifying experimental design. To demonstrate their use, we show their application in multiplexed western blotting. The sensitivity of Qdot conjugates is found to be superior to fluorescent dyes, and comparable to, or potentially better than, enhanced chemiluminescence. We show a true biological validation using a four-colour multiplexed western blot against a complex cell lysate background, and have significantly improved previously reported non-specific binding of the Qdots to cellular proteins.

Analytical procedures for the simultaneous detection of platinum analogues

DUE TO COPYRIGHT RESTRICTIONS ONLY AVAILABLE FOR CONSULTATION AT ASTON UNIVERSITY LIBRARY AND INFORMATION SERVICES WITH PRIOR ARRANGEMENT

Contrast discrimination with simultaneous monocular and dichoptic masks

In experiments reported elsewhere at this conference, we have revealed two striking results concerning binocular interactions in a masking paradigm. First, at low mask contrasts, a dichoptic masking grating produces a small facilitatory effect on the detection of a similar test grating. Second, the psychometric slope for dichoptic masking starts high (Weibull ß~4) at detection threshold, becomes low (ß~1.2) in the facilitatory region, and then unusually steep at high mask contrasts (ß~5.5). Neither of these results is consistent with Legge's (1984 Vision Research 24 385 - 394) model of binocular summation, but they are predicted by a two-stage gain control model in which interocular suppression precedes binocular summation. Here, we pose a further challenge for this model by using a 'twin-mask' paradigm (cf Foley, 1994 Journal of the Optical Society of America A 11 1710 - 1719). In 2AFC experiments, observers detected a patch of grating (1 cycle deg-1, 200 ms) presented to one eye in the presence of a pedestal in the same eye and a spatially identical mask in the other eye. The pedestal and mask contrasts varied independently, producing a two-dimensional masking space in which the orthogonal axes (10X10 contrasts) represent conventional dichoptic and monocular masking. The resulting surface (100 thresholds) confirmed and extended the observations above, and fixed the six parameters in the model, which fitted the data well. With no adjustment of parameters, the model described performance in a further experiment where mask and test were presented to both eyes. Moreover, in both model and data, binocular summation was greater than a factor of v2 at detection threshold. We conclude that this two-stage nonlinear model, with interocular suppression, gives a good account of early binocular processes in the perception of contrast. [Supported by EPSRC Grant Reference: GR/S74515/01]

Simultaneous interrogation of fiber Bragg grating sensors using an acoustooptic tunable filter

We describe a novel technique to provide demultiplexing of fiber Bragg grating sensors, interrogated using interferometric wavelength shift detection. Amplitude modulation of multiple radio frequency driving signals allows an acoustooptic tunable filter to provide wavelength demultiplexing. We demonstrated a noise limited strain resolution of 150 nanostrain/v(Hz) and a crosstalk better than -50 dB.

Simultaneous estimation of global background synaptic inhibition and excitation from membrane potential fluctuations in layer III neurons of the rat entorhinal cortex in vitro

It is becoming clear that the detection and integration of synaptic input and its conversion into an output signal in cortical neurons are strongly influenced by background synaptic activity or "noise." The majority of this noise results from the spontaneous release of synaptic transmitters, interacting with ligand-gated ion channels in the postsynaptic neuron [Berretta N, Jones RSG (1996); A comparison of spontaneous synaptic EPSCs in layer V and layer II neurones in the rat entorhinal cortex in vitro. J Neurophysiol 76:1089-1110; Jones RSG, Woodhall GL (2005) Background synaptic activity in rat entorhinal cortical neurons: differential control of transmitter release by presynaptic receptors. J Physiol 562:107-120; LoTurco JJ, Mody I, Kriegstein AR (1990) Differential activation of glutamate receptors by spontaneously released transmitter in slices of neocortex. Neurosci Lett 114:265-271; Otis TS, Staley KJ, Mody I (1991) Perpetual inhibitory activity in mammalian brain slices generated by spontaneous GABA release. Brain Res 545:142-150; Ropert N, Miles R, Korn H (1990) Characteristics of miniature inhibitory postsynaptic currents in CA1 pyramidal neurones of rat hippocampus. J Physiol 428:707-722; Salin PA, Prince DA (1996) Spontaneous GABAA receptor-mediated inhibitory currents in adult rat somatosensory cortex. J Neurophysiol 75:1573-1588; Staley KJ (1999) Quantal GABA release: noise or not? Nat Neurosci 2:494-495; Woodhall GL, Bailey SJ, Thompson SE, Evans DIP, Stacey AE, Jones RSG (2005) Fundamental differences in spontaneous synaptic inhibition between deep and superficial layers of the rat entorhinal cortex. Hippocampus 15:232-245]. The function of synaptic noise has been the subject of debate for some years, but there is increasing evidence that it modifies or controls neuronal excitability and, thus, the integrative properties of cortical neurons. In the present study we have investigated a novel approach [Rudolph M, Piwkowska Z, Badoual M, Bal T, Destexhe A (2004) A method to estimate synaptic conductances from membrane potential fluctuations. J Neurophysiol 91:2884-2896] to simultaneously quantify synaptic inhibitory and excitatory synaptic noise, together with postsynaptic excitability, in rat entorhinal cortical neurons in vitro. The results suggest that this is a viable and useful approach to the study of the function of synaptic noise in cortical networks. © 2007 IBRO.

Simultaneous measurement of temperature and external refractive index by use of a hybrid grating in D fiber with enhanced sensitivity by HF etching

We propose a dual-parameter optical sensor device achieved by UV inscription of a hybrid long-period grating-fiber Bragg grating structure in D fiber. The hybrid configuration permits the detection of the temperature from the latter's response and measurement of the external refractive index from the former's response. In addition, the host D fiber permits effective modification of the device's sensitivity by cladding etching. The grating sensor has been used to measure the concentrations of aqueous sugar solutions, demonstrating its potential capability to detect concentration changes as small as 0.01%.

Optical sensor based on hybrid LPG/FBG in D-fiber for simultaneous refractive index and temperature measurement

A dual-parameter optical sensor has been realized by UV-writing a long-period and a Bragg grating structure in D-fiber. The hybrid configuration permits the detection of the temperature from the latter and measuring the external refractive index from the former responses, respectively. The employment of the D-fiber allows as effective modification and enhancement of the device sensitivity by cladding etching. The grating sensor has been used to measure the concentrations of aqueous sugar solutions, demonstrating the potential capability to detect concentration changes as small as 0.01%.

Simultaneous measurement of temperature and external refractive index by use of a hybrid grating in D fiber with enhanced sensitivity by HF etching

We propose a dual-parameter optical sensor device achieved by UV inscription of a hybrid long-period grating-fiber Bragg grating structure in D fiber. The hybrid configuration permits the detection of the temperature from the latter's response and measurement of the external refractive index from the former's response. In addition, the host D fiber permits effective modification of the device's sensitivity by cladding etching. The grating sensor has been used to measure the concentrations of aqueous sugar solutions, demonstrating its potential capability to detect concentration changes as small as 0.01%.

Simultaneous strain and temperature measurement of advanced 3-D braided composite materials using an improved EFPI/FBG system

Simultaneous strain and temperature measurement for advanced 3-D braided composite materials using fibre-optic sensor technology is demonstrated, for the first time. These advanced 3-D braided composites can virtually eliminate the most serious problem of delamination for conventional composites. A tandem in-fibre Bragg-grating (FBG)/extrinsic Fabry-Perot interferometric sensor (EFPI) system with improved accuracy has been used to facilitate simultaneous temperature and strain measurement in this work. The non-symmetric distortion of the optical spectrum of the FBG, due to the combination of the FBG and the EFPI, is observed for the first time. Experimental and theoretical studies indicate that this type of distortion can affect the measurement accuracy seriously and it is mainly caused by the modulation of the periodic output of the EFPI. A simple method has been demonstrated to improve the accuracy for detection of the wavelength-shift of the FBG induced by temperature change. A strain accuracy of ∼ ±20 με and a temperature accuracy of ∼ ±1 °C have been achieved, which can meet the requirements for practical applications of 3-D braided composites. © 2002 Elsevier Science Ltd. All rights reserved.

Simultaneous interrogation of fiber Bragg grating sensors using an acoustooptic tunable filter

In this letter, we describe a novel technique to provide demultimplexing of fiber Bragg grating sensors, interrogated using interferometric wavelength shift detection. Amplitude modulation of multiple radio frequency driving signals allows au acoustooptic tunable filter to provide wavelength demultiplexing. We demonstrated a noise limited strain resolution of 150 nanostrain/√Hz and a crosstalk better than -50 dB.

Optical sensor based on hybrid LPG/FBG in D-fiber for simultaneous refractive index and temperature measurement

A dual-parameter optical sensor has been realized by UV-writing a long-period and a Bragg grating structure in D-fiber. The hybrid configuration permits the detection of the temperature from the latter and measuring the external refractive index from the former responses, respectively. The employment of the D-fiber allows as effective modification and enhancement of the device sensitivity by cladding etching. The grating sensor has been used to measure the concentrations of aqueous sugar solutions, demonstrating the potential capability to detect concentration changes as small as 0.01%.

Simultaneous measurement for liquid level and refractive index based on all-fiber modal interferometer

A simple fiber sensor capable of simultaneous measurement of liquid level and refractive index (RI) is proposed and experimentally demonstrated. The sensing head is an all-fiber modal interferometer manufactured by splicing an uncoated single-mode fiber with two short sections of multimode fiber. The interference pattern experiences blue shift along with an increase of axial strain and surrounding RI. Owing to the participation of multiple cladding modes with different sensitivities, the height and RI of the liquid could be simultaneously measured by monitoring two dips of the transmission spectrum. Experimental results show that the liquid level and RI sensitivities of the two dips are 245.7 pm/mm, -38 nm/RI unit (RIU), and 223.7 pm/mm, -62 nm/RIU, respectively. The approach has distinctive advantages of easy fabrication, low cost, and high sensitivity for liquid level detection with the capability of distinguishing the RI variation simultaneously. © 2013 Copyright Taylor and Francis Group, LLC.