Live cell imaging using wide-field microscopy and deconvolution.
The use of fluorescence imaging methods, most recently based on fluorescent protein technology, and the availability of high quality fluorescence imaging systems have driven a revolution in cell and molecular biology. Live cell imaging, especially using fluorescence, is now used in a wide variety of assays in academic and commercial laboratories. The use of this technology requires particular attention to be paid to cell engineering, the design of the image acquisition system, the imaging protocol, and subsequent processing and analytic methods. In this review, we discuss each of these steps, highlighting practical techniques developed by us and others.
Swedlow
Jason R
JR
Division of Gene Regulation and Expression, MSI/WTB Complex, University of Dundee, Dundee, Scotland. s.swedlow@dundee.ac.uk
Platani
Melpomeni
M
D000465:Algorithms; D000818:Animals; D018929:Cell Culture Techniques; D015194:Diffusion Chambers, Culture; D005057:Eukaryotic Cells / Q000166:cytology* / Q000378:metabolism; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence; D012815:Signal Processing, Computer-Assisted
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins