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 samples in the embedding machine for embedding. First, place the melted paraffin in the cassettes, and take out the tissue from the dehydration box before the paraffin is solidified, then put the cassettes and attach the corresponding label according to the requirements of 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.
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 liquid from Servicebio Inc, muscle tissue can choose environmental protection GD fixed from Servicebio Inc. We do not recommended acidic fixative solutions such as Bouin, which have an adverse effect on cell nuclear staining.
|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.