Framework for morphometric classification of cells in imaging flow cytometry

Imaging flow cytometry is an emerging technology that combines the statistical power of flow cytometry with spatial and quantitative morphology of digital microscopy. It allows high-throughput imaging of cells with good spatial resolution, while they are in flow. This paper proposes a general framework for the processing/classification of cells imaged using imaging flow cytometer. Each cell is localized by finding an accurate cell contour. Then, features reflecting cell size, circularity and complexity are extracted for the classification using SVM. Unlike the conventional iterative, semi-automatic segmentation algorithms such as active contour, we propose a noniterative, fully automatic graph-based cell localization. In order to evaluate the performance of the proposed framework, we have successfully classified unstained label-free leukaemia cell-lines MOLT, K562 and HL60 from video streams captured using custom fabricated cost-effective microfluidics-based imaging flow cytometer. The proposed system is a significant development in the direction of building a cost-effective cell analysis platform that would facilitate affordable mass screening camps looking cellular morphology for disease diagnosis. Lay description In this article, we propose a novel framework for processing the raw data generated using microfluidics based imaging flow cytometers. Microfluidics microscopy or microfluidics based imaging flow cytometry (mIFC) is a recent microscopy paradigm, that combines the statistical power of flow cytometry with spatial and quantitative morphology of digital microscopy, which allows us imaging cells while they are in flow. In comparison to the conventional slide-based imaging systems, mIFC is a nascent technology enabling high throughput imaging of cells and is yet to take the form of a clinical diagnostic tool. The proposed framework process the raw data generated by the mIFC systems. The framework incorporates several steps: beginning from pre-processing of the raw video frames to enhance the contents of the cell, localising the cell by a novel, fully automatic, non-iterative graph based algorithm, extraction of different quantitative morphological parameters and subsequent classification of cells. In order to evaluate the performance of the proposed framework, we have successfully classified unstained label-free leukaemia cell-lines MOLT, K562 and HL60 from video streams captured using cost-effective microfluidics based imaging flow cytometer. The cell lines of HL60, K562 and MOLT were obtained from ATCC (American Type Culture Collection) and are separately cultured in the lab. Thus, each culture contains cells from its own category alone and thereby provides the ground truth. Each cell is localised by finding a closed cell contour by defining a directed, weighted graph from the Canny edge images of the cell such that the closed contour lies along the shortest weighted path surrounding the centroid of the cell from a starting point on a good curve segment to an immediate endpoint. Once the cell is localised, morphological features reflecting size, shape and complexity of the cells are extracted and used to develop a support vector machine based classification system. We could classify the cell-lines with good accuracy and the results were quite consistent across different cross validation experiments. We hope that imaging flow cytometers equipped with the proposed framework for image processing would enable cost-effective, automated and reliable disease screening in over-loaded facilities, which cannot afford to hire skilled personnel in large numbers. Such platforms would potentially facilitate screening camps in low income group countries; thereby transforming the current health care paradigms by enabling rapid, automated diagnosis for diseases like cancer.

Computational study of viscous effects on lobed mixer flow features and performance

This article describes a computational study of viscous effects on lobed mixer flowfields. The computations, which were carried out using a compressible, three-dimensional, unstructured-mesh Navier-Stokes solver, were aimed at assessing the impacts on mixer performance of inlet boundary-layer thickness and boundary-layer separation within the lobe. The geometries analyzed represent a class of lobed mixer configurations used in turbofan engines. Parameters investigated included lobe penetration angles from 22 to 45 deg, stream-to-stream velocity ratios from 0.5 to 1.0, and two inlet boundary-layer displacement thicknesses. The results show quantitatively the increasing influence of viscous effects as lobe penetration angle is increased. It is shown that the simple estimate of shed circulation given by Skebe et al. (Experimental Investigation of Three-Dimensional Forced Mixer Lobe Flow Field, AIAA Paper 88-3785, July, 1988) can be extended even to situations in which the flow is separated, provided an effective mixer exit angle and height are defined. An examination of different loss sources is also carried out to illustrate the relative contributions of mixing loss and of boundary-layer viscous effects in cases of practical interest.

Generation of long, laminar plasma jets at atmospheric pressure and effects of flow turbulence

Long, laminar plasma jets at atmospheric pressure of pure argon and a mixture of argon and nitrogen with jet length up to 45 fi,Hes its diameter could be generated with a DC are torch by! restricting the movement of arc root in the torch channel. Effects of torch structure, gas feeding, and characteristics of power supply on the length of plasma jets were experimentally examined. Plasma jets of considerable length and excellent stability could be obtained by regulating the generating parameters, including are channel geometry gas flow I ate, and feeding methods, etc. Influence of flow turbulence at the torch,nozzle exit on the temperature distribution of plasma jets was numerically simulated. The analysis indicated that laminar flow plasma with very low initial turbulent kinetic energy will produce a long jet, with low axial temperature gradient. This kind of long laminar plasma jet could greatly improve the controllability for materials processing, compared with a short turbulent are let.

Preliminary Investigation of Fluid Flow in Meso-Scale Rectangular Tube

The fluid flow associated with micro and meso scale devices is currently of interest. Experiments were performed to study the fluid flow in meso-scale channels. A straight flow tube was fabricated with 1.0x4.0mm^2 in rectangular cross section and 200mm in length, which was made of quartz for flow visualization and PIV measurements. Reynolds numbers were ranged from 311 to over 3105. The corresponding pressure drop was from 0.65KPa to over 16.58KPa between the inlet and outlet of the tube. The micro PIV was developed to measure the velocity distribution in the tube. A set of microscope object lens was mounted ahead of CCD camera to obtain optimized optical magnification on the CCD chip. The velocity distributions near the outlet of the tube were measured to obtain full-developed flow. A CW laser beam was focused directly on the test section by a cylinder lens to form a small light sheet. Thus, high power density of light was formed on the view region. It is very important to the experiment while the velocity of the flow reaches to a few meters per second within millimeter scale. In this case, it is necessary to reduce exposure time to microseconds for PIV measurements. In the present paper, the experimental results are compared with the classical theories.

Dissimilar autogenous full penetration welding of superalloy K418 and 42CrMo steel by a high power CWNd : YAG laser

Experiments of autogenous laser full penetration welding between dissimilar cast Ni-based superalloy K418 and alloy steel 42CrMo flat plates with 3.5 mm thickness were conducted using a 3 kW continuous wave (CW) Nd:YAG laser. The influences of laser welding velocity, flow rate of side-blow shielding gas, defocusing distance were investigated. Microstructure of the welded seam was characterized by optical microscopy (OM), scanning electron microscopy (SEM) and X-ray diffraction (XRD) and energy dispersive spectrometer (EDS). Mechanical properties of the welded seam were evaluated by microhardness and tensile strength testing. Results show that high quality full penetration laser-welded joint can be obtained by optimizing the welding velocity, flow rate of shielding gas and defocusing distance. The laser-welded seam have non-equilibrium solidified microstructures consisting of gamma-FeCr0.29Ni0.16C0.06 austenite solid solution dendrites as the dominant and very small amount of super-fine dispersed Ni3Al gamma' phase and Laves particles as well as MC needle-like carbides distributed in the interdendritic regions. Although the microhardness of the laser-welded seam was lower than that of the base metal, the strength of the joint was equal to that of the base metal and the fracture mechanism showed fine ductility. (c) 2007 Elsevier B.V. All rights reserved.

The asymptotic near-tip solution for mode-iii crack in steady growth in power hardening media

Two local solutions, one perpendicular and one parallel to the direction of solar gravitational field, are discussed. The influence of gravity on the gas-dynamical process driven by the piston is discussed in terms of characteristic theory, and the flow field is given quantitatively. For a typical piston trajectory similar to the one for an eruptive prominence, the velocity of the shock front which locates ahead the transient front is nearly constant or slightly accelerated, and the width of the compressed flow region may be kept nearly constant or increased linearly, depending on the velocity distribution of the piston. Based on these results, the major features of the transient may be explained. Some of the fine structure of the transient is also shown, which may be compared in detail with observations.

The mechanical effects of water flow on fish eggs and larvae

The impact of mechanical stresses upon ichthyoplankton entrained in power plant cooling systems has long been considered negligible. Arguments and evidence exist, however, to show that such a supposition is not universally true, especially in nuclear power plants. The mechanisms of mechanical damage can be detailed in terms of pressure change, acceleration, and shear stress with in the fluid flow field. Laboratory efforts to quantify the effects of mechanical stress have been very sparse. A well-planned bioassay is urgently needed. (PDF has 11 pages.)

A numerical investigation on the flow physics of koi carp (Cyprinus carpio koi) burst-and-coast swimming

Abstract—Burst-and-coast is the most common locomotion type in freely routine swimming of koi carps (Cyprinus carpio koi), which consists of a burst phase and a coast phase in each cycle and mostly leads to a straight-line trajectory. Combining with the tracking experiment, the flow physics of koi carp’s burst-andcoast swimming is investigated using a novel integrated CFD method solving the body-fluid interaction problem. The dynamical equations of a deforming body are formulated. Following that, the loose-coupled equations of the body dynamics and the fluid dynamics are numerically solved with the integrated method. The two burst modes, MT (Multiple Tail-beat) and HT (Half Tail-beat), which have been reported by the experiments, are investigated by numerical simulations in this paper. The body kinematics is predicted and the flow physics is visualized, which are in good agreement with the corresponding experiments. Furthermore, the optimization on the energy cost and several critical control mechanisms in burst-and-coast swimming of koi carps are explored, by varying the parameters in its selfpropelled swimming. In this paper, energetics is measured by the two mechanical quantities, total output power CP and Froude efficiency Fr. Results and discussion show that from the standpoint of mechanical energy, burst-and-coast swimming does not actually save energy comparing with steady swimming at the same average speed, in that frequently changing of speed leads to decrease of efficiency.

Fluctuation characteristics of arc voltage and jet flow in a non-transferred dc plasma generated at reduced pressure

A torch with a set of inter-electrode inserts between the cathode and the anode/nozzle with a wide nozzle exit was designed to generate plasma jets at chamber pressures of 500–10 000 Pa. The variation of the arc voltage was examined with the change in working parameters such as gas flow rate and chamber pressure. The fluctuation in the arc voltage was recorded with an oscilloscope, and the plasma jet fluctuation near the torch exit was observed with a high-speed video camera and detected with a double-electrostatic probe. Results show that the 300 Hz wave originated from the tri-phase rectified power supply was always detected under all generating conditions. Helmholtz oscillations over 3000 Hz was detected superposed on the 300 Hz wave at gas flow rates higher than 8.8 slm with a peak to valley amplitude lower than 5% of the average voltage value. No appreciable voltage fluctuation caused by the irregular arc root movement is detected, and mechanisms for the arc voltage and jet flow fluctuations are discussed.