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发表于 2016-7-8 15:10:44
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本帖最后由 龙吟风 于 2016-7-8 15:13 编辑
Transmission electron microscopy.
The samples were dissolved in HEPES (4-[2-hydroxyethyl]-1-piperazine ethanesulfonic acid) buffer, and a drop of the suspension was placed on a sheet of parafilm.
A carbon-coated copper grid was floated on the drop for 10 seconds. Then, the grid was removed, and excess liquid was drained by touching the edge of the grid against a piece of clean filter paper. The grid was touched onto a drop of 2% uranyl acetate or phosphotungstic acid, pH 7.0, for approximately 5 seconds, and excess liquid was drained off. The grid was allowed to dry for several minutes and then examined using a JEM-1200 EX microscope (JEOL,Akishima, Japan) at 80 kiloelectron volts.
Tanaka,Y，H．Kamohara,K．Kinoshita,et a1．Clinical impact of serum exosomal microRNA-2 1 as a clinical biomarker in human esophageal squamouscell carcinoma．[J]Cancer,201 3．1 1 9(6)：P．1 1 59—67．该种方法运用在多篇文章中
Transmission electron microscopy
TEM was performed by the method of Thery et al. [50].Exosomes isolated by ExoQuick reagent were suspended in PBS. A drop of this solution was allowed to settle on a goldcoated grid, fixed in 1 % glutaraldehyde, washed for 2 min in double-distilled water, and incubated in uranyl oxylate for 5 min, followed by incubation with three separate drops of methyl cellulose with uranyl acetate—5 min with the first two drops and 10 min with the last drop—and finally, methyl cellulose-uranyl acetate was removed by slow-drag on edge on filter paper. Exosomes were visualized by standard TEM with a Philips CM120 microscope
Munagala, R., F. Aqil and R.C. Gupta, Exosomal miRNAs as biomarkers of recurrent lung cancer. Tumor Biology, 2016.以上就是用过的方法，电镜室的老师说步骤不详细，很多没法做。
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