Longitudinal in vivo imaging of adult Danionella cerebrum using standard confocal microscopy.
Longitudinal in vivo imaging of adult Danionella cerebrum using standard confocal microscopy.
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DOI：
10.1242/dmm.049753
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发表时间：
2022-12-01
影响因子：
4.3
通讯作者：
中科院分区：
文献类型：
--
作者：
大脑丹尼奥氏菌是一种新的脊椎动物模型，它提供了一个令人兴奋的机会来可视化完整成年动物的动态生物学过程。该模型的主要优点包括体积小、终身光学透明、遗传适宜性和遗传时间短。建立一种可靠的方法，在保持存活的同时，对成年D脑进行体内纵向成像，将允许对完整的活动物的发育、疾病发生和进展、伤口愈合和衰老等各种过程进行深入的基于图像的研究。在这里，描述了一种使用定制设计的3D打印成像室对成人D大脑进行延长和纵向共聚焦实时成像的方法。建立了两个转基因D脑细胞系以测试成像系统，即TG(mpeg1：Dendra2)和TG(kdrl：mCherry-CAAX)。第一条线用于显示巨噬细胞和小胶质细胞，第二条线用于空间配准。通过使用这种方法，在几天的过程中，观察到在动态平衡以及对刺伤或双光子诱导的脑损伤的反应中，免疫细胞的形态和行为的差异。透明成年脊椎动物模型丹尼奥拉大脑的体内和纵向共聚焦成像实验装置、材料和程序。
Danionella cerebrum is a new vertebrate model that offers an exciting opportunity to visualize dynamic biological processes in intact adult animals. Key advantages of this model include its small size, life-long optical transparency, genetic amenability and short generation time. Establishing a reliable method for longitudinal in vivo imaging of adult D. cerebrum while maintaining viability will allow in-depth image-based studies of various processes involved in development, disease onset and progression, wound healing, and aging in an intact live animal. Here, a method for both prolonged and longitudinal confocal live imaging of adult D. cerebrum using custom-designed and 3D-printed imaging chambers is described. Two transgenic D. cerebrum lines were created to test the imaging system, i.e. Tg(mpeg1:dendra2) and Tg(kdrl:mCherry-caax). The first line was used to visualize macrophages and microglia, and the second for spatial registration. By using this approach, differences in immune cell morphology and behavior during homeostasis as well as in response to a stab wound or two-photon-induced brain injury were observed in intact adult fish over the course of several days. Experimental imaging setup, materials and procedures for in vivo and longitudinal confocal imaging of the transparent adult vertebrate model Danionella cerebrum.
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影响因子：
3.1
作者：
Harvie, Elizabeth A.;Green, Julie M.;Huttenlocher, Anna
通讯作者：
Huttenlocher, Anna
影响因子：
4.6
作者：
Jung, Hyun Min;Castranova, Daniel;Weinstein, Brant M.
通讯作者：
Weinstein, Brant M.
影响因子：
3.5
作者：
Kobat, Demirhan;Horton, Nicholas G.;Xu, Chris
通讯作者：
Xu, Chris
影响因子：
1.2
作者：
Crilly, Siobhan;Njegic, Alexandra;Kasher, Paul R.
通讯作者：
Kasher, Paul R.
影响因子：
1.5
作者：
Britz, Ralf;Conway, Kevin W.
通讯作者：
Conway, Kevin W.