果蝇成虫盘的培养与活体成像

Cultivation and Live Imaging of Drosophila Imaginal Discs.

作者信息

Restrepo Simon, Zartman Jeremiah J, Basler Konrad

机构信息

Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, Zurich, CH-8057, Switzerland.

Department of Chemical and Biomolecular Engineering, University of Notre Dame, 182 Fitzpatrick Hall, Notre Dame, IN, 46556, USA.

出版信息

Methods Mol Biol. 2016;1478:203-213. doi: 10.1007/978-1-4939-6371-3_11.

DOI:10.1007/978-1-4939-6371-3_11

PMID:27730583

Abstract

The ex vivo cultivation and live imaging of wing discs open exciting new research avenues by overcoming the limitations of end-point analysis of fixed tissues. Here we describe how to prepare an optimized wing disc culture medium (WM1) and how to dissect and arrange wing discs for cultivation and live imaging. This protocol enables the study of dynamic phenomena such as cell division and delamination as well as the use of pharmacological compounds and biosensors. Wing discs cultured and imaged as described here, maintain constant levels of proliferation during the first ten hours of culture.

摘要

通过克服固定组织终点分析的局限性，翅芽的体外培养和实时成像开辟了令人兴奋的新研究途径。在这里，我们描述了如何制备优化的翅芽培养基（WM1），以及如何解剖和布置翅芽用于培养和实时成像。该方案能够研究细胞分裂和分层等动态现象，以及使用药理化合物和生物传感器。按照此处所述进行培养和成像的翅芽，在培养的前十个小时内保持恒定的增殖水平。