| Co-Investigator(Kenkyū-buntansha) |
KAWANA Akio NTT Basic Research Laboratories, Material Science Research Laboratory, Kawana Re, 物質科学研究部, 川名研究グループリー
HOMMA Tomoo Tokyo University of Agriculture and Technology Faculty of Technology, Research A, 工学部, 助手 (80242246)
NEMOTO Yasuyuki Tokyo University of Agriculture and Technology, Faculty of Technology, Lecturer, 工学部, 講師 (70202249)
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| Research Abstract |
Microbioassay for stress signal to living cells has recently been recognized important from the viewpoint new drug development and clinical analysis. This study aimed to develop an analyzing system of cellular responses to multiple stress signals. Multi-channnel sub-micron capillary (MMC) was used to apply a chemical or electrical stress signal to a particular plant cell and, at the same time, to detect the response signal of the cell or another cell. The response signals were chemicel, electrical, or electrochemical.
Anionic fluorescence dye, lucifer yellow (LY), was introduced into a cell of cultured tobacco (BY-2, derived from Nicotiana tabacum L.cv.Bright Yellow-2) or a leaf cell of Tradescantia virginiana L.as an example of a chemical signal. The diffusion pattern of LY was found a useful indicator to confirm the tip position of the capillary. In the same way, H^+, K^+, Cl were injected in a cell and its injection was ascertained with respective ion-selective microelectrodes which were devised with MMC.The reaction of enzymes such as luciferase and urease which were injected in a cell was also measured in vivo.
In order to estimate an exact voltage acting between the intercellular membrane, a high performence impedance analyzing system was developed using a patch-clamp amplifier and a lock-in amplifier. The membrane impedance of a intercellular membrane of BY-2 was, for example, determined as a parallel circuit of 2-7G OMEGA and 500pF.
The present system has enabled the application of multiple stress signals separately or simultaneously, or repeatedly to the same single cell. This is a marked advantageous point in studying complicated relationships between stress signal intensity and cellilar response intensity.
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