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The cryo-thin sectioning technique is an electron microscopy sample preparation technique developed to study structures and functions closer to the living state of the living organism. It is a thin slice after the sample is frozen and fixed, and the long-term fixation, dehydration and burial of the classic ultra-thin section method are omitted. The sample excitation can be well preserved before and after the slice without strong chemical treatment; especially the small soluble components will be extracted, making the electron microscope image closer to the living state of the organism. It can be used in cell immunochemistry, self-development of soluble components, especially in X-ray micro-regional component analysis. Frozen ultra-thin slicer operation: It can be carried out by adding a high-pressure freezer to the ultra-thin slicer . The preparation procedure for frozen ultrathin sections can be of two types:
1. No solution preparation method: including several steps such as material extraction, rapid cold tracking, freezing replacement , slicing, drying, dyeing or non-staining, microscopic examination, and the like. It is mainly used for radio white development and X-ray micro-analysis of soluble substances.
2. Fixed sample preparation method: including taking materials, aldehydes, sample embedding or not embedding, cryoprotection IR, rapid freezing section, dyeing and microscopic examination. Suitable for morphological, cytochemical and immunocytochemical studies.
After obtaining the training, the user can use the ultra-thin slicer by himself. The steps are as follows:
1) Boot
2) Check if it is in the slicing mode and check if the top light is normal.
3) Put the embedding block into the appropriate collet and tighten with a hex wrench to fix the embedding block; (then install the collet on the repair station for repair);
4) Put the chuck into the arm and screw the screw up by hand (no hex wrench);
5) Install the glass knife/diamond knife on the knife table, screw the screw by hand to fix the knife, and pay attention to check whether the knife table is fixed on the base;
6) Check the knife clearance angle, adjust the turret angle, select the appropriate chuck angle, etc.;
7) Set the slicing window (upper and lower ranges);
8) the knife;
9) Select the slice speed and thickness used;
10) Fill the sipe with water, pay attention to the smooth surface;
11) Slice and slice;
12) Shut down, clean the slicer, take away personal items, and keep the sliced ​​table clean and tidy;
13) Register the usage record.