1. Types of tissue and cell specimens: (1) Tissue specimens: 1. Paraffin sections: preserved tissue morphology 2. Frozen sections: preserved antigenicity (2) Cell specimens: 1. Tissue prints 2. Cell culture slices (Cell slide) 3. Cell smear 2. Tissue sampling 1. Brain tissue and nerve tissue should be fixed by perfusion and then fixed. 2. Precautions for tissue selection: (1) Fresh specimens: tissues require immersion in fixative solution within 30min ~ 2h after separation, especially in molecular biology. Animal tissue requires that the heart be collected while the heart is still beating. (2) Pay attention to prevent the influence of human factors. Knife and shear should be fast and avoid squeezing. (3) The thickness of the tissue block is 0.1 ~ 0.3cm, and the size can be determined according to needs. 3. Preparation method of live cell specimens (1) After the cells are cultured in large quantities, the cells are made into a suspension by digestion (106) and coated on a slide coated with a slice adhesive, and then dried and fixed. (2) Culture the cells directly on the coverslip. (3) The cells are directly cultured on 6-well or 9-well plates, and IHC is feasible after fixation. 4. Preparation of paraffin sections 1. Since paraffin is insoluble in water and alcohol reagents, it is only dissolved in xylene reagents. Therefore, the tissue contains a large amount of water after being fixed and washed, and needs to be dehydrated with ethanol reagents. After the water is removed, the tissue contains a large amount of ethanol. It must also be replaced with xylene that can dissolve the ethanol. The cells are weighed by a large number of paraffin branches, so that the tissue can be used for paraffin sections. 2. Small animal tissue dehydration, transparent and wax dipping time 75% ethanol 30min, 85% ethanol 30min, 95% ethanol â… 1h, â…¡ 1h, â…¢ overnight or 2h; anhydrous ethanol â… , â…¡ and â…¢ 30min each; xylene â… 15min, â…¡ 10min , â…¢ 10min; paraffin â… 30min, paraffin â…¡ 30min, paraffin â…¢ 1 ~ 2h. 5. Storage and use of frozen slices 1. After frozen slices are fixed and stored at -80 ° C for later use. 2. Frozen slices are taken out from -80 ° C. Immunohistochemical labeling can be carried out directly after being dried or dried by hair dryer. 1. Types of tissue and cell specimens: (1) Tissue specimens: 1. Paraffin sections: preserved tissue morphology 2. Frozen sections: preserved antigenicity (2) Cell specimens: 1. Tissue prints 2. Cell culture slices (Cell slide) 3. Cell smear 2. Tissue sampling 1. Brain tissue and nerve tissue should be fixed by perfusion and then fixed. 2. Precautions for tissue selection: (1) Fresh specimens: tissues require immersion in fixative solution within 30min ~ 2h after separation, especially in molecular biology. Animal tissue requires that the heart be collected while the heart is still beating. (2) Pay attention to prevent the influence of human factors. Knife and shear should be fast and avoid squeezing. (3) The thickness of the tissue block is 0.1 ~ 0.3cm, and the size can be determined according to needs. 3. Preparation method of live cell specimens (1) After the cells are cultured in large quantities, the cells are made into a suspension by digestion (106) and coated on a slide coated with a slice adhesive, and then dried and fixed. (2) Culture the cells directly on the coverslip. (3) The cells are directly cultured on 6-well or 9-well plates, and IHC is feasible after fixation. 4. Preparation of paraffin sections 1. Since paraffin is insoluble in water and alcohol reagents, it is only dissolved in xylene reagents. Therefore, the tissue contains a large amount of water after being fixed and washed, and needs to be dehydrated with ethanol reagents. After the water is removed, the tissue contains a large amount of ethanol. It must also be replaced with xylene that can dissolve the ethanol. The cells are weighed by a large number of paraffin branches, so that the tissue can be used for paraffin sections. 2. Small animal tissue dehydration, transparent and wax dipping time 75% ethanol 30min, 85% ethanol 30min, 95% ethanol â… 1h, â…¡ 1h, â…¢ overnight or 2h; anhydrous ethanol â… , â…¡ and â…¢ 30min each; xylene â… 15min, â…¡ 10min , â…¢ 10min; paraffin â… 30min, paraffin â…¡ 30min, paraffin â…¢ 1 ~ 2h. 5. Storage and use of frozen slices 1. After frozen slices are fixed and stored at -80 ° C for later use. 2. Frozen slices are taken out from -80 ° C. Immunohistochemical labeling can be carried out directly after being dried or dried by hair dryer. Source: Biotechnology Forum
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