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Introduction. Comparing different emitter classes and rationally designing the next generation of molecular and nanoscale probes for bioimaging applications require accurate and quantitative methods for the measurement of the key parameter photoluminescence quantum yield f.1 f equals the number of emitted per number of absorbed photons. This is particularly relevant for increasingly used fluorescence imaging in the short wave-infrared region (SWIR) ≥ 900 nm providing deeper penetration depths, a better image resolution, and an improved signal-to-noise or tumor-to-background ratio.2, 3 However, spectroscopic measurements in the SWIR are more challenging and require specific calibrations and standards.
For fluorescence microscopy, there is an increasing need for suitable calibration tools and reference materials for microscope calibration, the determination of performance parameters, and the regular control and validation of instrument performance. This is addressed in the BmWk-financed project FluMikal (WIPANO program) by two research groups from academia and two companies, that is coordinated by BAM. Here we present different approaches to liquid and solid fluorescence standards for the determination of the wavelength-dependent spectral sensitivity of fluorescence microscopes and ideas concerning the choice of suitable fluorescence lifetime standard for increasingly utilized fluorescence lifetime imaging (FLIM).
Standards for fluorescence analysis: From steady state fluorometry to fluorescence microscopy
(2006)
Spektrale Fluoreszenzstandards zur rückführbaren Charakterisierung von Fluoreszenzmesssystemen
(2004)