原始 paper： http://ajpregu.physiology.org/cgi/content/full/291/5/R1499 节录其中一段 Bioelectrical Measurements The intestinal flat sheets from discrete intestinal fragments were collected and prepared as described. Epithelia were lightly stretched to minimize the unstirred boundary layer, pinned over the circular aperture (1 cm2) of the chamber with the perimeter area lightly greased to minimize edge damage, and mounted in a recirculation vertical Ussing chamber (4-ml half-chamber volume). Measurement of bioelectrical variables was performed only in symmetric conditions ^^^^^^^^^ by using the basolateral saline composition described in Table 2 under cm2) was automatically calculated from voltage deflections after bipolar 200-ms pulses of 50 muA were injected every minute. Rt and short-circuit current ^^^^^^^^^^^^^^^^^^^^^ (Isc; muA/cm) were monitored by means of Ag-AgCl electrodes connected to the chambers by 3-mm bore agar bridges (3 M KCl in 3% agar). Clamping of epithelia to 0 mV and recording of variables ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ was performed using a microcomputer-controlled voltage-current Clamp Electronic with automatic correction for fluid resistance and electrode voltage asymmetry (KMSCI, Aachen, Germany).voltage clamp to 0 mV. Ep\ ithelial resistance Rt ( 这一段是在讲要怎麽测钙离子在海水硬骨鱼 肠道细胞从 apical 到 basal lateral 的 infux 和 efflux 有几点不太懂： 1.就实验设计的部份来说，为什麽要分 symmetric 跟 asymmetric condition? aymmetric 指的是 "maintaining the basolateral saline constant and changing the composition of the apical saline" 2.为什麽要测 short-circuit current？代表什麽意义？ 3."Clamping of epithelia to 0 mV"，patch clamp 不是针对单一culture後的细胞 做的吗？如果直接拿 fresh 的 tissue，也可以做喔？？how？ --

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