Method for spreading mouse spermatocyte chromosome

The invention discloses a method for spreading a mouse spermatocyte chromosome. According to the method, a mouse testicle is grinded, filtered, and centrifugally processed; a NaCl solution is taken asa hypotonic solution; a PBS mixed solution of paraformaldehyde and sodium dodecyl sulfate is taken a...

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Hauptverfasser: HUANG SHUNGUO, BU WENJIE, HE YU, YU YING, ZHU LEI, NIE LIUWANG, ZHU XIAOLEI, WANG PING, LI TIECHEN, WEI HUIJUN, GONG LEI, LIN AIQIN, WU ZHIHAO, GAO JIGUANG, YANG JIANKE
Format: Patent
Sprache:chi ; eng
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Zusammenfassung:The invention discloses a method for spreading a mouse spermatocyte chromosome. According to the method, a mouse testicle is grinded, filtered, and centrifugally processed; a NaCl solution is taken asa hypotonic solution; a PBS mixed solution of paraformaldehyde and sodium dodecyl sulfate is taken as a fixing solution to fix mouse spermatocyte on a glass slide; then a mouse SYCP3 monoclonal antibody is taken as a primary antibody; Dylight 549 fluorescence labeled goat-anti-mouse IgG second antibody is taken as a second antibody; a laser confocal microscopy is adopted to observe the spreadingof mouse spermatocyte, and the mitotic phase is recorded at the same time. The method can observe the morphological characteristics of all chromosomes more clearly, and promotes the research and analysis on the meiosis process of spermatocyte and pathogenesis of related diseases. 本发明公开了种用于小鼠精母细胞染色体铺展的方法,将小鼠睾丸破碎处理后并过滤离心后,以NaCl溶液作为低渗液;以多聚甲醛和十二烷基硫酸钠的PBS混合液作为固定液将小鼠精母细胞固定于玻片上;然后以小鼠SYCP3单克隆抗体为抗,Dylight 549荧光标记的山羊抗小鼠IgG二抗为二抗,采用激光