Bonaccorsi Silvia, Giansanti Maria G, Cenci Giovanni, Gatti Maurizio
Cold Spring Harb Protoc. 2012 Aug 1;2012(8):pdb.prot067355. doi: 10.1101/pdb.prot067355.
This protocol describes formaldehyde fixation of Drosophila testes. The procedure preserves chromosome morphology very well, allowing a clear visualization of the chromosome condensation-decondensation cycle. It also results in excellent preservation of microtubules for immunostaining. The main disadvantage of this fixation technique is its poor preservation of cell morphology, as viewed by phase-contrast optics. In addition, because it involves a rather hard squashing of formaldehyde-fixed testes, it results in meiotic figures which are flatter and larger than those obtained using methanol-acetone or paraformaldehyde fixation protocols.
本方案描述了果蝇睾丸的甲醛固定方法。该程序能很好地保存染色体形态，可清晰观察染色体的凝聚-解凝聚周期。它还能出色地保存微管以便进行免疫染色。这种固定技术的主要缺点是，在相差光学显微镜下观察时，细胞形态保存不佳。此外，由于该方法需要对甲醛固定的睾丸进行相当用力的压片，导致减数分裂图像比使用甲醇-丙酮或多聚甲醛固定方案得到的图像更扁平、更大。
