Preservation of blood and tissue samples for stable-carbon and stable-nitrogen isotope analysis
Preservation of blood and tissue samples for stable-carbon and stable-nitrogen isotope analysis
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DOI：
10.1139/z97-799
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发表时间：
1997-10-01
期刊：
影响因子：
--
通讯作者：
Gloutney, ML
中科院分区：
文献类型：
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作者：
Hobson, KA;Gibbs, HL;Gloutney, ML
从事收集动物材料进行稳定碳和稳定氮同位素分析的研究人员经常面临在运输到实验室之前需要保存组织的问题。在许多情况下，冷冻在现场是不可能的，因此我们研究了几种用于保存组织的技术的潜力。我们还包括用于DNA分析的保存技术，以评估它们如何改变组织中的δ（13）C和δ（15）N值，并最终评估存档的DNA样本是否可用于稳定同位素测定。组织包括鹌鹑（Coturnix coturnix japonica）和绵羊（Ovis aries）的血液和胸肌。血液的保存技术包括冷冻干燥（对照）、在预燃玻璃纤维滤纸上干燥以及储存在70%乙醇、10%缓冲福尔马林、ABI裂解缓冲液和Queen's裂解缓冲液中。8周后，使用裂解缓冲液和福尔马林导致血液中C-13和N-15的显著消耗。在玻璃纤维滤纸上干燥或在70%乙醇中储存的样品的值与对照品的值无显著差异。将肌肉组织冷冻干燥（对照）或储存在70%乙醇、10%缓冲福尔马林或DMSO溶液中。与对照相比，DMSO和福尔马林处理均导致C-13和N-15的显著消耗。只有70%乙醇处理没有导致肌肉中同位素比率的变化。如果无法冷冻，我们建议将血样干燥或储存在70%乙醇中。我们的研究提供了同位素校正因子的估计，可应用于DNA分析存档或保存在福尔马林中的组织。
Researchers engaged in collecting animal material for stable-carbon and stable-nitrogen isotope analysis are frequently faced with the need to preserve tissues prior to transportation to the laboratory. In many cases, freezing is not possible in the field, so we investigated the potential of several techniques for preserving tissues for this purpose. We also included preservation techniques used for DNA analyses in order to evaluate how they might alter delta(13)C and delta(15)N values in tissues and, ultimately, whether archived DNA samples could be used for stable-isotope assay. Tissues included blood and pectoral muscle from quail (Coturnix coturnix japonica) and blood from sheep (Ovis aries). Preservation techniques for blood included freeze-drying (control), drying on precombusted glass-fibre filter paper, and storing in 70% ethanol, 10% buffered formalin, ABI lysis buffer, and Queen's lysis buffer. After 8 weeks, the use of both lysis buffers and formalin resulted in significant depletion of C-13 and N-15 in blood. Values for samples dried on glass-fibre filter paper or stored in 70% ethanol did not differ significantly from those for the control. Muscle tissue was freeze-dried (control) or stored in 70% ethanol, 10% buffered formalin, or DMSO solution. Both the DMSO and formalin treatments resulted in significant depletion of C-13 and N-15 compared with the control. Only the 70% ethanol treatment did not result in changes to either isotope ratio in muscle. Where freezing is not possible, we recommend that blood samples be dried or stored in 70% ethanol. Our study provides an estimate of isotopic correction factors that may be applied to tissues archived for DNA analysis or stored in formalin.