Guiding Principles for Live Cell Imaging of Plants Using Confocal Microscopy
Live cell imaging using confocal microscope is an important and popular technique used by biologists to observe and understand biological events occuring in a living cell. It allows simultaneous understanding of the dynamics and functions of many cellular processes in living cells. The principles of live cell imaging are different from that of fixed cell imaging as in live cells, pigments and fluorescent biomolecules are present in their functional state unlike in fixed cell imaging where they are removed. This poses various challenges in live cell imaging, such as maintaining cell viability under high photobleaching conditions and the use of optimal fluorescent components to overcome the artifacts. Given the multitude of advantages of live cell imaging over conventional microscopy, the purpose of this chapter is to provide a basic understanding of the approaches used to visualize plant cells using confocal microscopy, discuss some common challenges encountered during live cell imaging and provide suggestions to overcome them.
KeywordsArabidopsis thaliana Confocal microscope Live cell imaging Plant cell Root Yellow fluorescent protein
MAS is supported by the Natural Sciences and Engineering Research Council of Canada funding. SS is supported by the Eyes High International Doctoral Scholarship and a Global Open Doctoral Scholarship from the University of Calgary. We thank Muhammad Jamshed for his technical assistance with the confocal microscope.
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