Department of Biochemistry, University of Washington, Seattle, WA 98105, USA; Program in Molecular and Cellular Biology, University of Washington School of Medicine, Seattle, WA 98105, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98105, USA.
Department of Biochemistry, University of Washington, Seattle, WA 98105, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98105, USA.
STAR Protoc. 2024 Mar 15;5(1):102895. doi: 10.1016/j.xpro.2024.102895. Epub 2024 Feb 16.
Functional studies in post-embryonic Xenopus tadpoles are challenging because embryonic perturbations often lead to developmental consequences, such as lethality. Here, we describe a high-throughput protocol for tail vein injection to introduce fluorescent tracers into tadpoles, which we have previously used to effectively inject morpholinos and molecular antagonists. We describe steps for safely positioning tadpoles onto agarose double-coated plates, draining media, injecting into the ventral tail vein, rehydrating plates, and sorting tadpoles by fluorescence with minimal injury for high-throughput experiments. For complete details on the use and execution of this protocol, please refer to Kakebeen et al., Patel et al., and Patel et al..
成体非洲爪蟾幼体的功能研究具有挑战性，因为胚胎干扰通常会导致发育后果，例如致死。在这里，我们描述了一种用于尾静脉注射的高通量方案，可将荧光示踪剂引入幼体中，我们之前曾使用该方案有效地注射了形态发生素和分子拮抗剂。我们描述了将幼体安全地放置在琼脂糖双层板上、排空培养基、向腹侧尾静脉注射、重新水化平板以及通过荧光对幼体进行分类的步骤，以实现高通量实验的最小损伤。有关此方案的使用和执行的完整详细信息，请参考 Kakebeen 等人、Patel 等人和 Patel 等人的研究。
