O'Neil J J, Sanford R L, Wasserman S, Tierney D F
J Appl Physiol Respir Environ Exerc Physiol. 1977 Nov;43(5):902-6. doi: 10.1152/jappl.1977.43.5.902.
We compared several sets of conditions used to estimate metabolism in rat lung slices. 14CO2 production from [14C]glucose, oxygen consumption, lactate production, and glucose consumption were used as measures of metabolic activity. The calculated results differed when we used 1) different techniques for estimating tissue weight, 2) tissue slices of 0.3-, 0.5-, 0.7-, and 1.0-mm thickness, 3) 95% air or 95% oxygen with 5% CO2 4) a delay after slice preparation and 4 degrees C and room temperature or periods of anoxia before incubation, 5) shaking rates of 60, 90, 120, or 150/min, 6) phosphate or bicarbonate buffers. Conditions of maximal activity were found using 95% O2 with 1.0-mm tissue slices, shaking at 120/min in phosphate buffer without periods of hypoxia or undue delays before incubation. Tissue weight should be obtained without exposure to aqueous solutions or dehydration by contact with cotton gauze or filter paper.
我们比较了用于评估大鼠肺切片代谢的几组条件。以[¹⁴C]葡萄糖产生¹⁴CO₂、耗氧量、乳酸生成量和葡萄糖消耗量作为代谢活性的指标。当我们使用以下条件时,计算结果有所不同:1)不同的组织重量估算技术;2)厚度为0.3毫米、0.5毫米、0.7毫米和1.0毫米的组织切片;3)95%空气或含5% CO₂的95%氧气;4)切片制备后的延迟时间以及4℃和室温条件,或孵育前的缺氧时间;5)60、90、120或150次/分钟的振荡速率;6)磷酸盐或碳酸氢盐缓冲液。在含1.0毫米组织切片的95% O₂条件下,于磷酸盐缓冲液中以120次/分钟振荡,且孵育前无缺氧期或过长延迟时间,可获得最大活性条件。获取组织重量时,应避免使其接触水溶液或因与纱布或滤纸接触而脱水。