Borgs P, Way D L, Witte M H, Witte C L
Department of Surgery, University of Arizona College of Medicine, Tucson 85724.
Alcohol. 1993 Jan-Feb;10(1):31-5. doi: 10.1016/0741-8329(93)90050-x.
Underestimation of ethanol (EtOH) volatility in vitro is a potential source of experimental error. EtOH (0-5% in culture medium) was added to 24- or 96-well tissue culture plates with standard low evaporation lids and incubated at 37 degrees C in humidified 7.5% CO2 and 92.5% air. After 72 hours, approximately 70% of the initial EtOH had disappeared from the aqueous phase of the plate (EtOH volatilization). EtOH concentrations gradually decreased in high-concentration wells (1-5%) and increased in low-concentration wells (0-0.1%) over time. This temporal redistribution of EtOH (EtOH diffusion) was detected after only 1 hour of incubation. Parafilm, Blenderm surgical tape, and ELISA plate-sealing tape barriers inconsistently or inadequately prevented EtOH volatilization and diffusion, but a newly designed plate-sealing clamp (PSC) apparatus inhibited this phenomenon. Rat hepatic sinusoidal endothelial cells cultured with the PSC apparatus maintained intact cell membranes for 72 hours and stable levels of monolayer permeability for at least 48 hours. By stabilizing in vitro EtOH concentrations, the PSC apparatus eliminates a potential source of major experimental error.
体外乙醇（EtOH）挥发性的低估是实验误差的一个潜在来源。将乙醇（培养基中含量为0 - 5%）添加到带有标准低蒸发盖的24孔或96孔组织培养板中，并在37℃、湿度为7.5%二氧化碳和92.5%空气的环境中孵育。72小时后，板中液相中约70%的初始乙醇消失（乙醇挥发）。随着时间推移，高浓度孔（1 - 5%）中的乙醇浓度逐渐降低，低浓度孔（0 - 0.1%）中的乙醇浓度升高。仅孵育1小时后就检测到了乙醇的这种时间性重新分布（乙醇扩散）。石蜡膜、Blenderm手术胶带和酶联免疫吸附测定板密封胶带屏障对乙醇挥发和扩散的阻止作用不一致或不充分，但一种新设计的板密封夹（PSC）装置可抑制这种现象。用PSC装置培养的大鼠肝窦内皮细胞在72小时内细胞膜保持完整，单层通透性水平至少在48小时内保持稳定。通过稳定体外乙醇浓度，PSC装置消除了一个主要的实验误差潜在来源。
