A Stochastic Model for Electron Multiplication Charge-Coupled Devices - From Theory to Practice

Electron multiplication charge-coupled devices (EMCCD) are widely used for photon counting experiments and measurements of low intensity light sources, and are extensively employed in biological fluorescence imaging applications. These devices have a complex statistical behaviour that is often not fully considered in the analysis of EMCCD data. Robust and optimal analysis of EMCCD images requires an understanding of their noise properties, in particular to exploit fully the advantages of Bayesian and maximum-likelihood analysis techniques, whose value is increasingly recognised in biological imaging for obtaining robust quantitative measurements from challenging data. To improve our own EMCCD analysis and as an effort to aid that of the wider bioimaging community, we present, explain and discuss a detailed physical model for EMCCD noise properties, giving a likelihood function for image counts in each pixel for a given incident intensity, and we explain how to measure the parameters for this model from various calibration images. © 2013 Hirsch et al.

Thermal imaging study of scalar transport in shallow wakes

The thermal imaging technique relies on the usage of infrared signal to detect the temperature field. Using temperature as a flow tracer, thermography is used to investigate the scalar transport in the shallow-water wake generated by an emergent circular cylinder. Thermal imaging is demonstrated to be a good quantitative flow visualization technique for studying turbulent mixing phenomena in shallow waters. A key advantage of the thermal imaging method over other scalar measurement techniques, such as the Laser Induced Fluorescence (LIF) and Planar Concentration Analysis (PCA) methods, is that it involves a very simple experimental setup. The dispersion characteristics captured with this technique are found to be similar to past studies with traditional measurement techniques. © 2012 Publishing House for Journal of Hydrodynamics.

Measuring EGFR Separations on Cells with ∼10 nm Resolution via Fluorophore Localization Imaging with Photobleaching

Detecting receptor dimerisation and other forms of clustering on the cell surface depends on methods capable of determining protein-protein separations with high resolution in the ∼10-50 nm range. However, this distance range poses a significant challenge because it is too large for fluorescence resonance energy transfer and contains distances too small for all other techniques capable of high-resolution in cells. Here we have adapted the technique of fluorophore localisation imaging with photobleaching to measure inter-receptor separations in the cellular environment. Using the epidermal growth factor receptor, a key cancer target molecule, we demonstrate ∼10 nm resolution while continuously covering the range of ∼10-80 nm. By labelling the receptor on cells expressing low receptor numbers with a fluorescent antagonist we have found inter-receptor separations all the way up from 8 nm to 59 nm. Our data are consistent with epidermal growth factor receptors being able to form homo-polymers of at least 10 receptors in the absence of activating ligands. © 2013 Needham et al.

A quality-guided displacement tracking algorithm for ultrasonic elasticity imaging.

Displacement estimation is a key step in the evaluation of tissue elasticity by quasistatic strain imaging. An efficient approach may incorporate a tracking strategy whereby each estimate is initially obtained from its neighbours' displacements and then refined through a localized search. This increases the accuracy and reduces the computational expense compared with exhaustive search. However, simple tracking strategies fail when the target displacement map exhibits complex structure. For example, there may be discontinuities and regions of indeterminate displacement caused by decorrelation between the pre- and post-deformation radio frequency (RF) echo signals. This paper introduces a novel displacement tracking algorithm, with a search strategy guided by a data quality indicator. Comparisons with existing methods show that the proposed algorithm is more robust when the displacement distribution is challenging.

Toxicity and imaging of multi-walled carbon nanotubes in human macrophage cells.

Multi-walled carbon nanotubes (MWNTs) have been proposed for use in many applications and concerns about their potential effect on human health have led to the interest in understanding the interactions between MWNTs and human cells. One important technique is the visualisation of the intracellular distribution of MWNTs. We exposed human macrophage cells to unpurified MWNTs and found that a decrease in cell viability was correlated with uptake of MWNTs due to mainly necrosis. Cells treated with purified MWNTs and the main contaminant Fe(2)O(3) itself yielded toxicity only from the nanotubes and not from the Fe(2)O(3). We used 3-D dark-field scanning transmission electron microscopy (DF-STEM) tomography of freeze-dried whole cells as well as confocal and scanning electron microscopy (SEM) to image the cellular uptake and distribution of unpurified MWNTs. We observed that unpurified MWNTs entered the cell both actively and passively frequently inserting through the plasma membrane into the cytoplasm and the nucleus. These suggest that MWNTs may cause incomplete phagocytosis or mechanically pierce through the plasma membrane and result in oxidative stress and cell death.

Curvature and wrinkling of premixed flame Kernels comparison between planar laser induced fluorescence (PLIF) and 3D direct numerical simulations (DNS)

A direct comparison between time resolved PLIF measurements of OH and two dimensional slices from a full three dimensional DNS data set of turbulent premixed flame kernels in lean methane/air mixture was presented. The local flame structure and the degree of flame wrinkling were examined in response to differing turbulence intensities and turbulent Reynolds numbers. Simulations were performed using the SEGA DNS code, which is based on the solution of the compressible Navier Stokes, species, and energy equations for a lean hydrocarbon mixture. For the OH PLIF measurements, a cluster of four Nd:YAG laser was fired sequentially at high repetition rates and used to pump a dye laser. The frequency doubled laser beam was formed into a sheet of 40 mm height using a cylindrical telescope. The combination of PLIF and DNS has been demonstrated as a powerful tool for flame analysis. This research will form the basis for the development of sub-grid-scale (SGS) models for LES of lean-premixed combustion systems such as gas turbines. This is an abstract of a paper presented at the 30th International Symposium on Combustion (Chicago, IL 7/25-30/2004).