Preparation method of transmission electron microscope sample with small number of cells
A technology for transmission electron microscope samples and a small number of cells, which is applied in the field of transmission electron microscope sample preparation of a small number of cells, and can solve the problems of easy loss and large number of cells required
Pending Publication Date: 2021-11-12
NANTONG UNIVERSITY
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[0004] The purpose of the present invention is to solve the defects in the prior art that the cell needs too much and is easy to lose
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[0030] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with specific examples.
[0031] In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention can also be implemented in other ways than described here. Therefore, the present invention is not limited to the specific embodiments disclosed below. limit.
[0032] A transmission electron microscope sample preparation method of a small amount of cells, comprising the following steps:
[0033] S1: Cell preparation:
[0034] Specifically, the cells cultured in a culture dish or a culture bottle are rinsed with PBS. In one embodiment, the pH value of the PBS is 7.2, and the number of rinses is 3 times, and each rinse time is 0.5 min; After the cells are added in the EP tube of 1.5ml, in one embodiment, the cells ar...
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Abstract
The invention provides a preparation method of a transmission electron microscope sample with a small number of cells, and relates to the technical field of transmission electron microscopes, and the preparation method comprises the following steps: adding cells into an EP tube, adding the collected cells into a precooled glutaraldehyde stationary liquid, centrifuging the EP tube, taking out a supernatant, adding 1% agarose gel, picking up a cell block mass and wrapping the cell block mass in the agarose gel; after cooling to a fixed state, cutting, adding glutaraldehyde stationary liquid again, cleaning by using a sodium dimethylformamide buffer solution, and dehydrating step by step by using ethanol; soaking the cells; gradually heating and polymerizing the cell sample to form an embedding block; polishing and cutting a cell dense block area in the embedding block into small resin blocks; and cutting into semi-thin slices, dyeing and positioning, grinding to only remain a target area, and cutting into ultra-thin slices by using a diamond cutter by taking the target area as the center to obtain the transmission electron microscope sample. According to the preparation method, the number of times of centrifugation is reduced, so that the change of cell morphology is avoided, and the real morphology of substances in the cells can be truly reflected.
Description
technical field [0001] The invention relates to the technical field of transmission electron microscopy, in particular to a method for preparing a sample for transmission electron microscopy with a small amount of cells. Background technique [0002] Transmission Electron Microscopy (TEM) is a very important tool and means for observing microstructure in the IC industry. Many critical dimensions of online detection need to be compared with the measured values of transmission electron microscopy, which is also an important way to solve many problems of online detection. [0003] Conventional transmission electron microscopy embedding methods have high requirements on the number of cell samples, and the method of centrifuging and collecting sediment for sample processing and embedding is basically adopted. However, too many times of centrifugation will cause changes in the shape of the tangible substance of the sample, and the operation steps are cumbersome and time-consu...
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IPC IPC(8): G01N23/04G01N1/30G01N1/36G01N1/28
CPCG01N23/04G01N1/36G01N1/28G01N2223/03G01N2223/102G01N2223/612
Inventor 刘芳李贵才徐佳炜王莹洁朱昌来张天一杨宇民
Owner NANTONG UNIVERSITY