Preparation method of frozen ultrathin slices

Abstract

The invention discloses a preparation method of frozen ultrathin slices. The method comprises the following steps: cutting a tissue sample by using a vibration slicer to obtain thin slices with the thickness of not more than 200 mu m; putting the thin slices into a sample table for high-pressure freezing; transferring the sample table loaded with the thin slices to a frozen ultrathin slicing machine, and pre-thinning the sample table loaded with the thin slices; transferring the pre-thinned sample table into a scanning transmission electron microscope; preparing a frozen ultrathin slice by using the frozen focused ion beam. Therefore, the thickness of the frozen ultrathin section prepared by the method is about 200nm and does not exceed 300nm, and the requirement of scanning transmission electron microscope imaging can be well met.

Description

technical field

[0001] The invention relates to the field of slice preparation, in particular to a method for preparing frozen ultrathin slices. Background technique

[0002] Frozen ultrathin sectioning is an electron microscope sample preparation technique that processes frozen samples into thin slices with a thickness of nanometers, so that they can be imaged on a transmission electron microscope. For biological samples, the hydrated samples obtained by rapid freezing without any chemical fixation can preserve the physiological state of the sample to the greatest extent and can be used for in situ structure research. However, if such a hydrated sample is ultra-thinly sliced ​​with an ordinary diamond knife, there will inevitably be problems such as compression of the sample along the cutting direction, serious knife marks on the slice surface, difficult operation and difficult collection of slices, etc., which cannot be widely used in the follow-up. Imaging and data colle...

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