The paraffin-embedding and section are most widely used in morphological research. It is mainly used to observe the morphological structure and judge pathological changes of tissues&cell. We provide a complete set of service such as paraffin embedding, section and staining etc., which includes sample selection, fixation, dehydration, transparent, paraffining, embedding, section and staining.
1.Tissue Selection: Fresh tissue should be fixed in fixative solution for more than 24h. After the tissue is taken out of the fixative solution,trim the target area with a scalpel in a fume hood, then place the trimmed tissue and corresponding label in the dehydration box.
2. Dehydration and paraffin infiltration:
100% alcohol I--0.5h
100% alcohol II--0.5h
Alcohol and Xylene mix--5 to 10 min
Xylene I--5 to 10 min
Xylene II--5 to 10 min
Paraffin (65°C) I--1h
Paraffin (65°C) II--1h
Paraffin (65°C) III--1h
3.Embedding: Put the tissue in the tissue embedding machine for embedding. First, place melted paraffin in the cassettes, and take out the tissue from the dehydration box before paraffin is solidified, then put the cassettes and attach the corresponding label according to the requirements of the embedding surface. Cool in a freezer at a temperature of -20 °C. After paraffin solidification, remove the paraffin block from the cassettes and trim accordingly.
4.Section: Place the trimmed paraffin block on a paraffin slicer and do section ,the thickness is about 3~4μm. The slices were floated on on a 40 °C warm water in a slice spreader, flatten the
tissue and pick up with a microscope slide , and baked in a 60 ° C oven. After water and paraffin was baked, taken out for storage at room temperature.
1. Fresh tissue should collect quickly: after body died, cut the target tissue quickly and put into the fixed solution. When cutting the tissue, the forceps should be avoided to press hardly, and the drawn knife should be used to drag the tissue back and forth.
2. Fresh tissue should be accurate: as accurately as possible to observe the target area .
3. Tissue material size: Normally the thickness of fresh tissue should not exceed 1cm while cutting.
4. Fixed container and fixed solution volume: the tissue is completely immersed in the fixed liquid, and the inverted container can move freely in the fixed liquid without sticking to the wall.
5. Fixed solution choice: animal tissue usually choose 4% paraformaldehyde, plant tissue usually choose FAA fixed solution, eyeball tissue may give preference to choose FAS eyeball fixed solution from Servicebio Inc, muscle tissue can choose environmental protection GD fixed solution from Servicebio Inc. We don’t recommended acidic fixative solutions such as Bouin, which have an adverse effect on cell nuclear staining.
Servicebio use MAS-GP type slide which is applied for IHC,IF and some special staining. It can prevent tissue fall from slide very well.