Experimental study of diffusion-based extraction from a cell suspension

A recently proposed application of microfluidics is the post-thaw processing of biological cells. Numerical simulations suggest that diffusion-based extraction of the cryoprotective agent dimethyl sulfoxide (DMSO) from blood cells is viable and more efficient than centrifugation, the conventional method of DMSO removal. In order to validate the theoretical model used in these simulations, a prototype was built and the flow of two parallel streams, a suspension of Jurkat cells containing DMSO and a wash stream that contained neither cells nor DMSO, was characterized experimentally. DMSO transport in a rectangular channel (depth 500 μm, width 25 mm and overall length 125 mm) was studied as a function of three dimensionless parameters: depth ratio of the streams, cell volume fraction in the cell solution, and the Peclet number (Pe) based on channel depth, average flow rate and the diffusion coefficient for DMSO in water. In our studies, values of Pe ranged from O(10³) to O(10⁴). Laminar flow was ensured by keeping the Reynolds number between O(1) and O(10). Experimental results based on visual and quantitative data demonstrate conclusively that a microfluidic device can effectively remove DMSO from liquid and cell laden streams without compromising cell recovery. Also, flow conditions in the microfluidic device appear to have no adverse effect on cell viability at the outlet. Further, the results demonstrate that we can predict the amount of DMSO removed from a given device with the theoretical model mentioned previously.     

Oxidation of proteins: Basic principles and perspectives for blood proteomics

Protein oxidation mechanisms result in a wide array of modifications, from backbone cleavage or protein crosslinking to more subtle modifications such as side chain oxidations. Protein oxidation occurs as part of normal regulatory processes, as a defence mechanism against oxidative stress, or as a deleterious processes when antioxidant defences are overcome. Because blood is continually exposed to reactive oxygen and nitrogen species, blood proteomics should inherently adopt redox proteomic strategies. In this review, we recall the biochemical basis of protein oxidation, review the proteomic methodologies applied to analyse redox modifications, and highlight some physiological and in vitro responses to oxidative stress of various blood components.     

A high-voltage cardiac stimulator for field shocks of a whole heart in a bath

Defibrillators are a critical tool for treating heart disease; however, the mechanisms by which they halt fibrillation are still not fully understood and are the subject of ongoing research. Clinical defibrillators do not provide the precise control of shock timing, duration, and voltage or other features needed for detailed scientific inquiry, and there are few, if any, commercially available units designed for research applications. For this reason, we have developed a high-voltage, programmable, capacitive-discharge stimulator optimized to deliver defibrillation shocks with precise timing and voltage control to an isolated animal heart, either in air or in a bath. This stimulator is capable of delivering voltages of up to 500 V and energies of nearly 100 J with timing accuracy of a few microseconds and with rise and fall times of 5 micros or less and is controlled only by two external timing pulses and a control computer that sets the stimulation parameters via a LABVIEW interface. Most importantly, the stimulator has circuits to protect the high-voltage circuitry and the operator from programming and input-output errors. This device has been tested and used successfully in field shock experiments on rabbit hearts as well as other protocols requiring high voltage.     

High precision electrostatic potential calculations for cylindrically symmetric lenses

A method is developed for a potential calculation within cylindrically symmetric electrostatic lenses using mesh relaxation techniques, and it is capable of considerably higher accuracies than currently available. The method involves (i) creating very high order algorithms (orders of 6, 8, and 10) for determining the potentials at points in the net using surrounding point values, (ii) eliminating the effect of the large errors caused by singular points, and (iii) reducing gradients in the high gradient regions of the geometry, thereby allowing the algorithms used in these regions to achieve greater precisions-(ii) and (iii) achieved by the use of telescopic multiregions. In addition, an algorithm for points one unit from a metal surface is developed, allowing general mesh point algorithms to be used in these situations, thereby taking advantage of the enhanced precision of the latter. A maximum error function dependent on a sixth order gradient of the potential is defined. With this the single point algorithmic errors are able to be viewed over the entire net. Finally, it is demonstrated that by utilizing the above concepts and procedures, the potential of a point in a reasonably high gradient region of a test geometry can realize a precision of less than 10(-10).     

Practical considerations for high spatial and temporal resolution dynamic transmission electron microscopy

Although recent years have seen significant advances in the spatial resolution possible in the transmission electron microscope (TEM), the temporal resolution of most microscopes is limited to video rate at best. This lack of temporal resolution means that our understanding of dynamic processes in materials is extremely limited. High temporal resolution in the TEM can be achieved, however, by replacing the normal thermionic or field emission source with a photoemission source. In this case the temporal resolution is limited only by the ability to create a short pulse of photoexcited electrons in the source, and this can be as short as a few femtoseconds. The operation of the photo-emission source and the control of the subsequent pulse of electrons (containing as many as 5 x 10(7) electrons) create significant challenges for a standard microscope column that is designed to operate with a single electron in the column at any one time. In this paper, the generation and control of electron pulses in the TEM to obtain a temporal resolution <10(-6)s will be described and the effect of the pulse duration and current density on the spatial resolution of the instrument will be examined. The potential of these levels of temporal and spatial resolution for the study of dynamic materials processes will also be discussed.     

Polarized light imaging of white matter architecture

Polarized light imaging (PLI) is a method to image fiber orientation in gross histological brain sections based on the birefringent properties of the myelin sheaths. The method uses the transmission of polarized light to quantitatively estimate the fiber orientation and inclination angles at every point of the imaged section. Multiple sections can be assembled into a 3D volume, from which the 3D extent of fiber tracts can be extracted. This article describes the physical principles of PLI and describes two major applications of the method: the imaging of white matter orientation of the rat brain and the generation of fiber orientation maps of the human brain in white and gray matter. The strengths and weaknesses of the method are set out.     

Mild body cooling impairs attention via distraction from skin cooling

Many contemporary workers are routinely exposed to mild cold stress, which may compromise mental function and lead to accidents. A study investigated the effect of mild body cooling of 1.0 degree C rectal temperature (Tre) on vigilance (i.e. sustained attention) and the orienting of spatial attention (i.e. spatially selective processing of visual information). Vigilance and spatial attention tests were administered to 14 healthy males and six females at four stages (pre-immersion, deltaTre = 0, -0.5 and - 1.0 degree C ) of a gradual, head-out immersion cooling session (18-25 deltaC water), and in four time-matched stages of a contrast session, in which participants sat in an empty tub and no cooling took place. In the spatial attention test, target discrimination times were similar for all stages of the contrast session, but increased significantly in the cooling phase upon immersion (deltaTre = 0 degrees C), with no further increases at deltaTre = -0.5 and - 1.0 degree C. Despite global response slowing, cooling did not affect the normal pattern of spatial orienting. In the vigilance test, the variability of detection time was adversely affected in the cooling but not the contrast trials: variability increased at immersion but did not increase further with additional cooling. These findings suggest that attentional impairments are more closely linked to the distracting effects of cold skin temperature than decreases in body core temperature.     

The utility of a virtual reality locomotion interface for studying gait behavior

OBJECTIVE: To investigate the effect of optic flow on gait behavior during treadmill walking using an immersive virtual reality (VR) setup and compare it with conventional treadmill walking (TW) and overground walking (OW). BACKGROUND: Previous research comparing TW with OW speculated that a lack of optic flow (relative visual movement between a walker and the environment) during TW may have led to perceptual cue conflicts, resulting in differences in gait behavior, as compared with OW. METHOD: Participants walked under three locomotion conditions (OW, TW, and TW with VR [TWVR]) under three walking constraint conditions (no constraint, a temporal/pacing constraint, and a spatial/path-following constraint). Presence questionnaires (PQs) were administered at the close of the TWVR trials. Trials were subjected to video analysis to determine spatiotemporal and kinematics variables used for comparison of locomotion conditions. RESULTS: ANOVA revealed gait behavior during TWVR to be between that of OW and TW. Speed and cadence during TWVR were significantly different from those of TW, whereas knee angle was comparable to that of OW. Correlation analysis of PQ scores with gait measures revealed a positive linear association of the distraction subfactor of the PQ with walking speed during TWVR, suggesting an increase in the sense of presence in the virtual environment led to increases in walking speed. CONCLUSION: The results demonstrate that providing optic flow during TW through VR has an impact on gait behavior. APPLICATION: This study provides a basis for developing simple VR locomotion interface setups for gait research.     

Surface dependent representations for illumination insensitive image comparison

We consider the problem of matching images to tell whether they come from the same scene viewed under different lighting conditions. We show that the surface characteristics determine the type of image comparison method that should be used. Previous work has shown the effectiveness of comparing the image gradient direction for surfaces with material properties that change rapidly in one direction. We show analytically that two other widely used methods, normalized correlation of small windows and comparison of multiscale oriented filters, essentially compute the same thing. Then, we show that for surfaces whose properties change more slowly, comparison of the output of whitening filters is most effective. This suggests that a combination of these strategies should be employed to compare general objects. We discuss indications that Gabor jets use such a mixed strategy effectively, and we propose a new mixed strategy. We validate our results on synthetic and real images