Bone and calcified tissue biopsies compound decalcification decalcification method of preparation and liquid.docVIP

 PAGE \* MERGEFORMAT 8 Bone and calcified tissue biopsies compound decalcification decalcification method of preparation and liquid [Keywords:] bone and calcified tissue; compound decalcification fluid; decalcification method; Improvement In addition to acid Bone and calcified tissue is very hard, be made of a few microns thin sectioning difficult. Commonly used method is the first decalcified, so that the organization become soft and then sliced, it is crucial decalcification technology. Decalcifying agent currently used mostly inorganic acid type agents, although the effect of decalcification better, but often causes tissue patterns, especially the destruction of cell structure and affect bone tissue morphology and research. To this end, we use the compound decalcification decalcification solution, not only able to maintain morphology smoothly and for special staining and slicing, are presented below: 1 Materials Clinical medical students on the dog OK after surgery, take tibia were sawn 1 cm long, and then split into four sections along the longitudinal tibia. With normal saline or 0.1 mol / L of PBS (pH7.2 ~ 7.4) buffer, washed two times, and then into the 3% glutaraldehyde and 4% paraformaldehyde fixed 1:1 mixture of 4 h; SHARP3A65-type microwave oven (Japan , maximum power 850 W, sub-5 file adjustable). Decalcification solution of Compound: Hydrochloric acid 8 ml, acetic acid 5 ml and salicylic acid 10 g; followed by adding 87 ml of distilled water mixed into the compound prepared decalcification solution. 2 Methods 2.1 decalcification method Plastic basket mounted specimens will be placed on the inclusion of medium-sized flat-bottomed beaker compound decalcification solution, and then place it in a large flat-bottomed beaker with ice water or a microwave dedicated plastic container into the microwave oven 2 file 10 min, remove the replacement by the ice demineralized water, refrigeration and heat-absorbing liquid used in cold water, set the fur