Biomedical subjects

Takanori Ichiki

Publications and source records attributed to Takanori Ichiki.

3 recordsLinked to original sources

PubMed2005

Measurement of the electrophoretic mobility of sheep erythrocytes using microcapillary chips.

Cell electrophoretic mobility (EPM) can be used to characterize individual cells. The purpose of this study is to establish reproducible and reliable cell EPM values obtained using microcapillary electrophoresis (microCE) chips. We studied cell electrophoresis on microCE chips through the comprehensive measurement of EPM and zeta potential. The inner wall of microchannels in microCE chips was coated with three kinds of reagents, namely bovine serum albumin (BSA), gelatin, and 2-methacryloyloxyethylphosphorylcholine (MPC) polymer to prevent nonspecific adhesion and interaction between cells and the inner wall. Electrophoresis was conducted in phosphate-buffered saline (pH 4-9) using erythrocytes extracted from sheep whole blood. Electroosmotic flow (EOF) mobility was measured using noncharged particles, and then the true EPM was calculated by subtracting the EOF mobility from the electromigration. MPC polymer coatings in microCE chips reduced the zeta potential of the inner wall and fully prevented nonspecific adhesion. EPM data obtained using microCE chips were almost the same and reproducible over a wide range of pH irrespective of the coating reagent used. In conclusion, reliability in the measurement of cell EPM using microCE chips was realized.

Animals↗

PubMed2004

Non-destructive on-chip cell sorting system with real-time microscopic image processing.

Studying cell functions for cellomics studies often requires the use of purified individual cells from mixtures of various kinds of cells. We have developed a new non-destructive on-chip cell sorting system for single cell based cultivation, by exploiting the advantage of microfluidics and electrostatic force. The system consists of the following two parts: a cell sorting chip made of poly-dimethylsiloxane (PDMS) on a 0.2-mm-thick glass slide, and an image analysis system with a phase-contrast/fluorescence microscope. The unique features of our system include (i) identification of a target from sample cells is achieved by comparison of the 0.2-microm-resolution phase-contrast and fluorescence images of cells in the microchannel every 1/30 s; (ii) non-destructive sorting of target cells in a laminar flow by application of electrostatic repulsion force for removing unrequited cells from the one laminar flow to the other; (iii) the use of agar gel for electrodes in order to minimize the effect on cells by electrochemical reactions of electrodes, and (iv) pre-filter, which was fabricated within the channel for removal of dust contained in a sample solution from tissue extracts. The sorting chip is capable of continuous operation and we have purified more than ten thousand cells for cultivation without damaging them. Our design has proved to be very efficient and suitable for the routine use in cell purification experiments.

Journal Article↗

PubMed2002

Immunoelectrophoresis of red blood cells performed on microcapillary chips.

Immunoanalysis of blood cells on a microcapillary electrophoresis (nuCE) chip has been studied using sheep erythrocytes (ShE) as an example. Two different buffer solutions, the phosphate-buffered saline (PBS) and the gelatin veronal buffer (GVB) were examined in regard to the electrokinetic transport behavior of ShE suspended in these solutions inside the rectangular channel engraved on a quartz chip. This clarified two advantages of the use of GVB for on-chip cell electrophoresis: gelatin coatings prevent (i) nonspecific sticking of ShE on the channel wall, and cause (ii) an appreciable reduction in the zeta potential of the wall suppressing the electroosmotic flow of the buffer solution. As a result ShE suspended in the GVB can smoothly migrate from the cathode to the anode, which is the opposite flow direction of immunoglobulin G (IgG) antibodies under the physiological pH condition of 7.4. Based on these results, on-chip capillary cell immunoelectrophoresis of ShE and rabbit anti ShE antibodies (IgG) have been proposed and successfully accomplished using the GVB. It is demonstrated that the variation of the cell migration velocity originating from the change in the surface charge after binding antibodies is applicable to the fast detection of immune reactions and also to single-cell typing.

Animals↗