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Preparation method of frozen ultrathin slices

A frozen ultra-thin section and thin section technology, which is applied in the preparation of test samples, sampling devices, instruments, etc., can solve the problem of unable to obtain images that can analyze tissue structure, avoid thinning time, ensure freezing quality, The effect of improving production efficiency

Pending Publication Date: 2020-07-14
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing frozen ultra-thin sections to solve the problem that the existing input-type cryofixation-FIB ultra-thin section-cryo-electron microscopy method cannot obtain images that can analyze tissue structures

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Embodiment Construction

[0049] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0050] According to an embodiment of the present invention, such as figure 1 As shown, a preparation method for frozen ultrathin sections is provided, comprising:

[0051] Step 101: Using a vibrating microtome to cut the tissue sample to obtain a thin section with a thickness not exceeding 200 μm.

[0052] The freezing depth of high-pressure freezing is much larger than that of rapid plunge freezing, and it can freeze biological tissue samples, but its depth is also...

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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...

Claims

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Application Information

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IPC IPC(8): G01N23/04G01N1/28G01N1/42G01N1/06H01J37/317
CPCG01N1/06G01N1/28G01N1/286G01N1/42G01N23/04G01N2001/2873H01J2237/31749
Inventor 孙磊张建国张丹阳季刚徐伟孙飞
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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