In vivo single-cell lineage tracing in zebrafish using high-resolution infrared laser-mediated gene induction microscopy

• DOI: 10.7554/elife.52024
• Publication Date: 2020-01-06T13:00:19Z
• Authors: Sicong He; Ye Tian; Shachuan Feng; Yi Wu; Xinwei Shen; Kani Chen; Yingzhu He; Qiqi Sun; Xuesong Li; Jin Xu; Zilong Wen; Jianan Y Qu
• Keywords: Fate mapping; Cell fate determination; Single-Cell Analysis; Cell type; Developmental Biology

Heterogeneity broadly exists in various cell types both during development and at homeostasis. Investigating heterogeneity is crucial for comprehensively understanding the complexity of ontogeny, dynamics, function specific types. Traditional bulk-labeling techniques are incompetent to dissect within population, while new single-cell lineage tracing methodologies invented last decade can hardly achieve high-fidelity labeling long-term in-vivo observation simultaneously. In this work, we developed a high-precision infrared laser-evoked gene operator heat-shock system, which uses laser-induced CreERT2 combined with loxP-DsRedx-loxP-GFP reporter precise tracing. vivo study indicated that system precisely label single brain, muscle hematopoietic zebrafish embryo. Using traced potential hemogenic endothelium (HE) posterior blood island (PBI) embryo found HEs PBI heterogeneous, contains least myeloid unipotent myeloid-lymphoid bipotent subtypes.