Multi-View Imaging of Drosophila Embryos

Multi-View Imaging;Microscope;Civil Engineering

Groh, Paul

University of Waterloo

关键词: Multi-View Imaging;    Microscope;    Civil Engineering;   

Others  :  https://uwspace.uwaterloo.ca/bitstream/10012/4024/1/Paul%20Groh%20Thesis.pdf

瑞士|英语

来源: UWSPACE Waterloo Institutional Repository

【 摘 要 】

There are several reasons for imaging a single, developing embryo from multiple view points. The embryo is a complex biomechanical system and morphogenesis movements in one region typicallyproduce motions in adjacent areas. Multi-view imaging can be used to observe morphogenesis and gain a better understanding of normal and abnormal embryo development. The system would allowthe embryo to be rotated to a specific vantage point so that a particular morphogenetic process may beobserved clearly. Moreover, a multi-view system can be used to gather images to create an accurate three-dimensional reconstruction of the embryo for computer simulations. The scope of this thesiswas to construct an apparatus that could capture multi-view images for these applications.A multi-view system for imaging live Drosophila melanogaster embryos, the first of its kind, is presented. Embryos for imaging are collected from genetically modified Drosophila stocks that contain a green fluorescing protein (GFP), which highlights only specific cell components. Theembryos are mounted on a wire that is rotated under computer control to desired viewpoints in front of the objective of a custom-built confocal microscope. The optical components for the orizontallyalignedmicroscope were researched, selected and installed specifically for this multi-viewingapparatus.The multiple images of the stacks from each viewpoint are deconvolved and collaged so as to show all of the cells visible from that view. The process of rotating and capturing images can be repeated for many angles over the course of one hour. Experiments were conducted to verify the repeatabilityof the rotation mechanism and to determine the number of image slices required to produce a satisfactory image collage from each viewpoint.Additional testing was conducted to establish that the system could capture a complete 360° view of the embryo, and a time-lapse study was done to verify that a developing embryo could be imagedrepeatedly from two separate angles during ventral furrow formation. An analysis of the effects of the imaging system on embryos in terms of photo-bleaching and viability is presented.