名稱 Bouin's Fixative / Bouin's 固定液
規(guī)格 100mL | 500mL
存儲 常溫避光保存,有效期12個月。
產(chǎn)品簡介
固定的目的在于保存細胞和組織的原有形態(tài)結(jié)構(gòu),固定劑能阻止內(nèi)源性溶酶體酶對自身組織和細胞的自溶、抑制細菌和霉菌的生長。固定劑通過凝固、生成添加化合物等使蛋白質(zhì)內(nèi)部結(jié)構(gòu)發(fā)生改變,從而使酶失活。固定液主要分為醛類固定液、汞類固定液、醇類固定液、氧化劑類固定液、苦味酸鹽類固定液等,較為常用的是醛類中的福爾馬林、醇類中的乙醇。
Phygene Bouin's固定液(Bouin's Fixative)主要由PA、甲醛、乙酸混合而成,PA可沉淀蛋白引起組織收縮,但不會使組織硬化, 其與甲醛和乙酸混合后,穿透速度加快,固定均勻,組織收縮輕微,對細胞的微細結(jié)構(gòu)顯示的很清晰,為一種良好的固定液。尤其對Masson三色法等的結(jié)締組織和肌纖維染色有媒染作用,經(jīng)其固定后的組織著色鮮艷。該固定液也可能軟化皮膚和肌腱,以利于切片。
使用參考
1、按實驗具體要求操作。
2、一般僅需固定30~60min。如果組織塊大,可適當延長固定時間,但不宜超過24h。
3、用Bouin固定液固定后,組織可稍微流水沖洗或不沖洗直接轉(zhuǎn)入70%的乙醇脫水,經(jīng)乙醇脫水時可除去大部分PA,組織留有一點黃色,對染色也無影響。
注意事項
1、Bouin's固定液對人體有一定的損害,請在通風好的環(huán)境下小心操作,避免吸入。
2、組織取材的厚度不同,固定時間也不同,對組織恰當?shù)倪x材有利于固定液的滲透。常規(guī)活檢組織比較適合的厚度為2~4 mm,一般不超過6 mm。
3、固定液的容量應(yīng)足夠,一般固定液與組織塊的體積比率應(yīng)大于10:1。如果容積不夠大,可以在固定期間更換1~3次固定液。
4、溫度對固定的影響很明顯,提高溫度可以加速固定作用,但溫度不宜過高。
5、取出新鮮組織后,應(yīng)及時固定,無法及時固定時,應(yīng)保存于生理鹽水中及時送檢。
Bouin solution, or Bouin's solution, is a compound fixative used in histology. It was invented by French biologist Pol Bouin and is composed of picric acid, acetic acid and formaldehyde in an aqueous solution. It is especially good for gastrointestinal tract biopsies because this fixative allows crisper and better nuclear staining than 10% neutral-buffered formalin. It is not a good fixative when tissue ultrastructure must be preserved for electron microscopy. However, it is a good fixative when tissue structure with a soft and delicate texture must be preserved. Formalin-fixed tissue is normally mordanted with Bouin solution for better staining results in the trichrome stains. The acetic acid in this fixative lyses red blood cells and dissolves small iron and calcium deposits in tissue. A variant in which the acetic acid is replaced with formic acid can be used for both fixation of tissue and decalcification.
The effects of the three chemicals in Bouin solution balance each other. Formalin causes cytoplasm to become basophilic but this effect is balanced by the effect of the picric acid. This results in excellent nuclear and cytoplasmic H&E staining. The tissue hardening effect of formalin is balanced by the soft tissue fixation of picric acid. The tissue swelling effect of acetic acid is balanced by the tissue shrinking effect of picric acid.
When using Bouin solution, several potential problems can arise. Due to the formalin in the solution, formalin pigment may be present when viewing tissue sections under the microscope. Wet tissue should be fixed in Bouin solution for less than 24 hours. Excess picric acid should be washed out of tissue using several alcohol and water solutions or staining quality may deteriorate over time. Wet tissue fixed in Bouin solution should be stored in an alcohol and water solution rather than Bouin solution. Since Bouin solution contains formaldehyde, picric acid and acetic acid, appropriate safety precautions for these substances should be taken and regulations followed. In particular, noting that picric acid can be explosive, sensitive to friction and shock when dry and in contact with some metals can form unstable metal picrates.
關(guān)鍵字: 4%多聚甲醛固定液
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