The paraffin-embedding and section are the most widely used methods in morphological research. It is mainly used to observe the morphological structure and pathological changes of tissues and cells. Our company provide a series of service about paraffin wax, section and staining, including sampling, fixation, dehydration, xylene transparent, paraffin wire, embedding, slicing and staining.
1.Tissue fixation:Dissect tissue as fast as possible, then immerse in fixative immediately. Trim tissue sample appropriately in chemical hood after fixation (at least 24 hours).
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: Process tissue sample in melted paraffin in cassettes. Adjust sample to desired orientation according to client’s request and label it. Freeze at -20°C until paraffin solidification complete. Take out paraffin block and trim accordingly.
1. Fresh tissue needs to be collected quickly: after the body dies, the target tissue needs to be cut quickly and put into the fixed solution. When cutting the tissue, the forceps should be avoided to press hard, and the drawn knife should be used to drag the tissue back and forth.
2. Fresh tissue materials should be accurate: as accurately as possible to the target area to be observed.
3. Tissue material size: generally, the thickness of fresh tissue should not exceed 1cm when 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 tissues usually choose 4% paraformaldehyde, plant tissue usually choose FAA fixed solution, eyeball tissues may give preference to choose FAS eyeball fixed liquid of Servicebio INC, muscle tissue can choose environmental protection GD fixed of Servicebio INC. We do not recommended acidic fixative solutions such as Bouin's , which have an effect on nuclear coloring.
|Coronal section||Horizontal section||Sagittal section|
|Cross section||Aortic valve/Ventricle||Atrium|
Intestine roll, observe the whole intestinal mucosa situation,highly recommend.
The whole intestine is in PFA fixation.
|Observe cornea,retina,optic disck,optic nerve|
|Zebra fish||Horizontal section||Sagittal section|
Tissue fixation provides for preservation of the tissue but leaves the tissue too pliable and without a means of support to allow for thin sections to be cut from the specimen.Ordinary tissue is recommended 4% paraformaldehyde(PFA) fixation solution. There is some requirement for sample preparation.
1.1 Select the special specimen bottle for specimen and make sure the tissue can move freely in the container.
1.2 Choose general tissue fixative solution which is more than 10 times volum of the tissue.
1.3 The thickness of tissue should not be more than 1cm and the area is not limited.
1.4 The embedding orientation and observation purpose should be clear.
Some special tissue need special tissue fixation solution to keep morphological integrity,such as eye ball,fat,plant,muscle.Servicebio is capable
offering those special fixation solution which is verified by massive researchers to be effective.
Regards to some special experiment such as Hybridization in Situ(ISH),TEM. Tissue also need to be preserved in special fixation solution.
Servicebio offer these kind of solution.