Though representing a major component of eukaryotic biodiversity, many microbial eukaryotes remain poorly studied, including the focus of the present work, testate amoebae of the order Arcellinida (Amoebozoa) and non-model lineages of ciliates (Alveolata). In particular, knowledge of genome structures and changes in genome content over the often-complex life cycles of these lineages remains enigmatic. However, the limited available knowledge suggests that microbial eukaryotes have the potential to challenge our textbook views on eukaryotic genomes and genome evolution. In this study, we developed protocols for DAPI (4′,6-diamidino-2-phenylindole) staining of Arcellinida nuclei and adapted protocols for ciliates. In addition, image analysis software was used to estimate the DNA content in the nuclei of Arcellinida and ciliates, and the measurements of target organisms were compared to those of well-known model organisms. The results demonstrate that the methods we have developed for nuclear staining in these lineages are effective and can be applied to other microbial eukaryotic groups by adjusting certain stages in the protocols.

尽管代表了真核生物多样性的一个主要组成部分，但许多微生物真核生物的研究仍然很少，包括目前工作的重点，Arcellinida 目 (Amoebozoa) 和纤毛虫的非模型谱系 (Alveolata)。特别是，在这些谱系通常复杂的生命周期中，关于基因组结构和基因组内容变化的知识仍然是个谜。然而，有限的可用知识表明微生物真核生物有可能挑战我们关于真核基因组和基因组进化的教科书观点。在这项研究中，我们开发了 Arcellinida 细胞核的 DAPI（4',6-二脒基-2-苯基吲哚）染色方案，并为纤毛虫制定了适应性方案。此外，图像分析软件用于估计Arcellinida和纤毛虫细胞核中的DNA含量， 结果表明，我们为这些谱系中的核染色开发的方法是有效的，并且可以通过调整协议中的某些阶段将其应用于其他微生物真核群体。