Image calibration in fluorescence microscopy
| Autor: | G. J. Brakenhoff, J. W. Hofstraat, G. J. Va N Rooij, Jurriaan M. Zwier |
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| Přispěvatelé: | Molecular Cytology (SILS, FNWI) |
| Rok vydání: | 2004 |
| Předmět: |
Histology
Microscope business.industry Chemistry Fluorescence Pathology and Forensic Medicine Characterization (materials science) law.invention Cell Line Mice Optics Microscopy Fluorescence law Light sheet fluorescence microscopy Microscopy Calibration Fluorescence microscope Animals Photoactivated localization microscopy business |
| Zdroj: | Journal of Microscopy, 216, 15-24. Wiley-Blackwell |
| ISSN: | 0022-2720 |
| Popis: | A fluorescence image calibration method is presented based on the use of standardized uniformly fluorescing reference layers. It is demonstrated to be effective for the correction of non-uniform imaging characteristics across the image (shading correction) as well as for relating fluorescence intensities between images taken with different microscopes or imaging conditions. The variation of the illumination intensity over the image can be determined on the basis of the uniform bleaching characteristics of the layers. This permits correction for the latter and makes bleach-rate-related imaging practical. The significant potential of these layers for calibration in quantitative fluorescence microscopy is illustrated with a series of applications. As the illumination and imaging properties of a microscope can be evaluated separately, the methods presented are also valuable for general microscope testing and characterization. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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