Accuracy and Precision in Quantitative Fluorescence Microscopy
| Title: | Accuracy and Precision in Quantitative Fluorescence Microscopy |
| Author: | Waters, Jennifer Catherine |
| Citation: | Waters, Jennifer C. 2009. Accuracy and precision in quantitative fluorescence microscopy. The Journal of Cell Biology 185(7): 1135-1148. |
| Full Text & Related Files: |
2712964.pdf (1.836Mb; PDF) 
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| Abstract: | The light microscope has long been used to document the localization of fluorescent molecules in cell biology research. With advances in digital cameras and the discovery and development of genetically encoded fluorophores, there has been a huge increase in the use of fluorescence microscopy to quantify spatial and temporal measurements of fluorescent molecules in biological specimens. Whether simply comparing the relative intensities of two fluorescent specimens, or using advanced techniques like Förster resonance energy transfer (FRET) or fluorescence recovery after photobleaching (FRAP), quantitation of fluorescence requires a thorough understanding of the limitations of and proper use of the different components of the imaging system. Here, I focus on the parameters of digital image acquisition that affect the accuracy and precision of quantitative fluorescence microscopy measurements. |
| Published Version: | doi:10.1083/jcb.200903097 |
| Other Sources: | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2712964/pdf/ |
| Terms of Use: | This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA |
| Citable link to this page: | http://nrs.harvard.edu/urn-3:HUL.InstRepos:4875892 |
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