脱蜡剂Histo-clearⅡ的实际应用举例

智能文字提取功能测试中

海德创业(北京)生物科技有限公司电话：010-525715025124812052571975；网址： http：//www.hedebio.com Email： hedebio@163.com海德创业(北京)生物科技有限公司1 小鼠组织的石蜡包埋和切片实验步骤举例： Paraffin Embedding and Sectioning of Tissues 1. Anesthetize a mouse using standard techniques (see Note 6). 2.. Perfuse the mouse (through the left ventricle of the heart) with fresh 10% formalin(prewarmed to 37°C). 3. Remove all the required tissues and fix in 10% formalin overnight.4.1The following day， wash the tissues twice for 20 min each time in DEPC-PBS. 5. Replace the PBS sequentially with 30， 50， and then 70% ethanol. Allow 30 min for eachincubation (with shaking). If using the brain， increase incubation times to I h. 6..FRemove the final ethanol wash and cover the tissue with fresh 70% ethanol. The tissuescan then be stored in 70% ethanol at-20°C. 7.To continue the processing， remove the 70% ethanol and incubate the tissues in 90%ethanol for 30 min (allow 1 h for the brain). Wash twice in 100% ethanol for 15 min eachtime (increase incubation time to 30 min for the brain). 8.Transfer the tissues to Histo-Clear II：ethanol(1：1) for 10 min， followed by 100% Histo-Clear II for 5 min. Do not incubate the tissues for more than 5 min in Histo-Clear II.because this makes them brittle when sectioning is carried out. .PFlace the tissues in a 1：1 mixture of wax：Histo-Clear II at 60°C for 10 min. 90. Transfer to a 3：1 mixture of wax：Histo-Clear II at 60°C for 10 min and finally to wax (at60°C) for about 30 min. Then place the tissues in fresh wax for a further 30 min. Whenworking with brain tissue， increase the wax treatment to at least 3 to 4 h. 11.U1sing plastic molds placed on a hot plate， embed the tissues in fresh wax. Ensure thatthere are no bubbles and that the wax is liquid when embedding the tissues； otherwise，thetissue blocks will give uneven sections. 12.Using a microtome， cut 10-um sections and remove creases in the ribbons of sections byfloating on top of water in a water bath at 37C. To place sections on slides， immerseSuperfrost slides in the water bath (close to a ribbon of sections) and use the slides to liftthe sections out. The sections will stick to the surface of the slides. Allow the slides to dryovernight at 37°C.