改良骨骼肌肌纤维分型ATP酶染色方法.docVIP

改良骨骼肌肌纤维分型ATP酶染色方法 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：改良骨骼肌肌纤维分型ATP酶染色方法 1 1 材料与方法 3 2 结　　果 4 3 讨　　论 5 文2：改良骨骼肌肌纤维分型ATP酶染色方法临床医学论文 6 1 材料与方法 8 2 结 果 9 3 讨 论 10 参考文摘引言： 11 原创性声明（模板） 12 文章致谢（模板） 13 正文 改良骨骼肌肌纤维分型ATP酶染色方法 文1：改良骨骼肌肌纤维分型ATP酶染色方法 ABSTRACT: Objective To study the effect of selfmodified Guth Samahas myosinATPase staining method on the classification of SD rat skeletal muscle fiber types. Methods 8μmthick cryostat sectio from rat chest muscles and biceps brachii muscles were trafected with cryostat. MyosinATPase staining was carried out according to the following procedure: ① Fixing sectio for 5min in fixative solution containing 40g/L paraformaldehyde; ② Riing slides in Trisrie solution and then preincubating them in alkaline preincubation solution for 15min; ③ Riing slides in Trisrie solution twice and then incubating them for 60min in incubation solution; ④ Washing slides for three times in 10g/L CaCl2 and then placing them in 20g/L CoCl2 for 3min; ⑤ Washing sides in distal water and then placing them in 10g/L (NH4)2S for 3min and; ⑥ Washing slides in running tap water for 3min， dehydrating in graded ethanol， clearing in xylene and mounting in neutral balsam. Results After being stained by modified myosinATPase staining method， both chest muscle and biceps brachii muscle samples from SD rats could be clearly identified as type Ⅰ fibe and type Ⅱfibe as the fibe of type Ⅰ were stained white while the fibe of type Ⅱ were stained brown. Conclusion Modified myosinATPase staining method is a simple and effective way for muscle fiber type classification and can be applied in skeletal muscle related study。 KEY WORDS: myosin ATPase; muscle fiber types; rat GuthSamaha肌球蛋白ATP酶染色骨骼肌纤维分型方法于上世纪70年代建立以来，已成为骨骼肌功能相关研究的重要手段之一[1]。该方法的应用极大地推动了骨骼肌相关研究的深入与发展。由于岁月的变迁，原GuthSamaha肌球蛋白ATP酶染色方案中所采用的特定缓冲溶液产品已停止生产，其配方也无从知晓，限制了该方法的推广应用。这或许是目前骨骼肌分型研究报道不多的原因之一。探寻更为简便、适用的肌球蛋白ATP酶染色方法对骨骼肌的相关研究具有现实意义。本研究采用自行改良的GuthSamaha肌球蛋白ATP酶染色方法