超低温保存白玉兰花粉细胞内游离钙离子的测定
首发时间:2009-01-16
摘要:以白玉兰花粉为材料,采用超纯水静置30min和花粉培养液水合30min两种预处理方式及常温(25℃)和低温(4℃)两种装载方法,将钙离子荧光探针Fluo-3, AM导入白玉兰花粉细胞。利用流式细胞仪检测了花粉细胞内游离钙离子的浓度变化。研究结果表明,白玉兰花粉经花粉培养液水合30min后,采用常温(25℃)装载Fluo-3, AM效果最好。超低温保存5秒引起白玉兰花粉细胞内钙离子浓度快速升高,而随着保存时间的延长钙离子浓度逐渐回落。
关键词: 白玉兰 花粉 钙离子 Fluo-3 AM 超低温保存
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The Measurement of Cytosolic Free Ca2+ in Magnolia denudata. Pollen Cells
Abstract:Magnolia denudate pollen were set in the ultrapure water for 30 minutes and in the culture media for the same time period as pretreatment. Fluorescent Fluo-3,AM was loaded into the cells under room temperature (25 oC) and low temperature (4 oC) to measure the fluorescence intensity of the intracellular Ca2+ by the flow cytometer. The research indicated that more Fluo-3, AM can be loaded when the pollen were pretreated by cultured media for 30 minutes and loaded under the room temperature (25oC). The cytosolic free Ca2+ of Magnolia denudate pollen increased sharply after cryopreserved for 5 seconds and gradually drops back with time passes by.
Keywords: Magnolia denudate pollen Ca2+ Fluo-3 AM cryopreservation
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