Gimlich R L, Braun J
Dev Biol. 1985 Jun;109(2):509-14. doi: 10.1016/0012-1606(85)90476-2.
In this note simple methods for the synthesis of several new fluorescent cell lineage tracers are described. These are fluorescent dextrans with average molecular weights of approximately 11 X 10(3), and with one or more fluorophore molecules covalently coupled to each dextran chain. These fluorescent dextrans are brighter than commercially obtainable products, and can be microinjected using either air-pressure injection or iontophoresis. They are long-lasting and have a uniform distribution in the cytoplasm of embryonic cells, clearly revealing very fine cell extensions such as cilia, axons, and filipodia. A method is also described for covalently attaching free amino groups to fluorescent dextran to make the tracers cofixable with cellular constituents by aldehyde treatment. Fluorescent dextran-amine tracers allow embryonic cell lineages to be studied in fixed, permeabilized, or sectioned embryos.
本笔记描述了几种新型荧光细胞谱系示踪剂的简单合成方法。这些是平均分子量约为11×10³的荧光葡聚糖，每个葡聚糖链上共价连接有一个或多个荧光团分子。这些荧光葡聚糖比市售产品更亮，可通过气压注射或离子电渗法进行显微注射。它们持久且在胚胎细胞的细胞质中分布均匀，能清晰显示非常细微的细胞突起，如纤毛、轴突和丝状伪足。还描述了一种将游离氨基共价连接到荧光葡聚糖上的方法，以使示踪剂通过醛处理与细胞成分共固定。荧光葡聚糖-胺示踪剂可用于在固定、通透或切片的胚胎中研究胚胎细胞谱系。
