A highly reproducible plant electrical signal–light-induced bioelectrogenesis (LIB) was obtained by means of periodic illumination/darkness stimulation of broad bean (Vicia faba L.) leaves. By stimulating the same position of the same leaf with different concentrations of NaCl, we observed that the amplitude and waveform of the LIB was correlated with the intensity of stimulation. This method allowed us to link dynamic ion fluxes induced by periodic illumination/darkness to salt stress. The self-referencing ion electrode technique was used to explore the ionic mechanisms of the LIB. Fluxes of H+, Ca2+, K+, and Cl− showed periodic changes under periodic illumination/darkness before and after 50 mM NaCl stimulation. Gray relational analysis was used to analyze correlations between each of these ions and LIB. The results showed that different ions are involved in surface potential changes at different stages under periodic illumination/darkness. The gray relational grade reflected the contribution of each ion to the change in surface potential at a certain time period. The ion fluxes data obtained under periodic illumination/darkness stimulation will contribute to the future development of a dynamic model for interpretation of electrophysiological events in plant cells.
Microscopic object recognition and analysis is very important in micromanipulation. Micromanipulation has been extensively used in many fields, e.g., micro-assembly operation, microsurgery, agriculture, and biological research. Conducting micro-object recognition in the in-situ measurement of tissue, e.g., in the ion flux measurement by moving an ion-selective microelectrode (ISME), is a complex problem. For living tissues growing at a rate, it remains a challenge to accurately recognize and locate an ISME to protect living tissues and to prevent an ISME from being damaged. Thus, we proposed a robust and fast recognition method based on local binary pattern (LBP) and Haar-like features fusion by training a cascade of classifiers using the gentle AdaBoost algorithm to recognize microscopic objects. Then, we could locate the electrode tip from the background with strong noise by using the Hough transform and edge extraction with an improved contour detection method. Finally, the method could be used to automatically and accurately calculate the relative distance between the two micro-objects in the microscopic image. The results show that the proposed method can achieve good performance in micro-object recognition with a recognition rate up to 99.14% and a tip recognition speed up to 14 frames/s at a resolution of 1360 × 1024. The max error of tip positioning is 6.10 μm, which meets the design requirements of the ISME system. Furthermore, this study provides an effective visual guidance method for micromanipulation, which can facilitate automated micromanipulation research.
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