Slice preparation method of non-decalcified osteochondral tissue and osteochondral tissue cell tracing and RNAScope combined test method

Abstract

The invention belongs to a biological detection technology, and particularly relates to a preparation method of a non-decalcified osteochondral tissue slice and an osteochondral tissue cell tracing and RNAScope combined test method. The problem that in the prior art, due to the fact that a non-decalcified osteochondral tissue slice is difficult to keep complete, the cell tracing and RNAScope combined testing technology cannot be applied to osteochondral tissue slices is solved. The technical scheme of the invention is as follows: the integrity of the osteochondral tissue during frozen sectionis guaranteed through a Kawamoto membrane, and then, a fluorescent cell tracing technology and an in-situ hybridization technology are combined to realize simultaneous observation of a cell tracing fluorescent signal and an RNAsocpe in-situ hybridization specific target RNA signal in the same osteochondral tissue section. The application range of the cell tracing and RNAScope combined test technology is expanded, and the application comprises (1) osteochondral tissue development research, (2) corresponding gene knockout efficiency analysis in osteochondral tissue, (3) research on influences ontranscription levels of other target genes after gene knockout, and (4) osteochondral tissue pathological analysis.

Description

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AI Technical Summary

Benefits of technology

This patented inventions aim to improve the accuracy and speed for studying biomolecules such as calcium (Ca), phosphorus (P), carbohydrates, nucleic acids, enzyme levels, oxidative stress reactions, inflammations caused by external factors like radiation exposure, chemical substances, pathogens, and changes overtime. By embedding these molecular structures into an ice crystal structure instead of freezing them afterwards, this technique allows scientists to examine how cells grow inside their own body while preserving its structural quality. Additionally, there are methods available where certain techniques may use oxygen gas laser irradiating onto the surface layer of calcified areas before making new ones. Overall, these technical improvements make better understanding and exploration of biomedical materials more efficient than current methods.

Problems solved by technology

This patents discusses how fluorochrome-based methods for studying bone growth processes (bone formation) involve combining certain molecules with an agent that specifically targets them during biological activity on the surface of living organisms such as cartilage. These agents may include enzyme catalysts like ribonucleases, lyases, etc., chemical compounds containing amino acids, peptides, proteins, nucleic acid bases, sugar residues, nucleators, antibodies, drugs, small metals, metal chelates, polymers, polysaccharide structures, lipocalites, dyes, cyanines, flavonoid pigments, phthalexine blue, green light absorbing materials, quantum dots, magnetic nanoparticles, iron oxidenanoferritans, calcium carbonate particles, silicon dioxane shell layers, silver nitrate solutions, polyethyleneimine films, and other substances known collectively called stainless steel wires.

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