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This optical absorption measurement of 4',6-Diamidino-2-phenylindole, [DAPI] were made by R.-C. A. Fuh in the summer of 1997 using a Cary 3. The absorption values were collected using a spectral bandwidth of 1.0 nm, a signal averaging time of 0.133 sec, a data interval of 0.25 nm, and a scan rate of 112.5 nm/min.
These measurements were scaled to make the molar extinction coefficient match the value of 27,000cm-1/M at 344.0nm (Haugland, 1992).
The fluorescence emission spectrum of 4',6-Diamidino-2-phenylindole, [DAPI] dissolved in water. The excitation wavelength was 350nm. The quantum yield of this molecule is 0.043 (HÃÂÃÂ¤rd, 1990). This spectrum was collected by in the summer of 1997 using a Spex FluoroMax. The excitation and emission monochromators were set at 1 mm, giving a spectral bandwidth of 4.25 nm. The data interval was 0.5 nm and the integration time was 2.0 sec.
Samples were prepared in 1cm pathlength quartz cells with absorbance less than 0.1 at the excitation and all emission wavelengths to uniformly illuminate across the sample, and to avoid the inner-filter effect. The dark counts were subtracted and the spectra were corrected for wavelength-dependent instrument sensitivity.
Dixon, J. M., M. Taniguchi and J. S. Lindsey (2005), "PhotochemCAD 2. A Refined Program with Accompanying Spectral Databases for Photochemical Calculations, Photochem. Photobiol., 81, 212-213.
Du, H., R.-C. A. Fuh, J. Li, L. A. Corkan and J. S. Lindsey (1998) PhotochemCAD: A computer-aided design and research tool in photochemistry. Photochem. Photobiol. 68, 141-142.
Haugland, R. P. (1992-1994) Molecular Probes. Handbook of Fluorescent Probes and Research Chemicals.
HÃÂÃÂ¤rd, T., P. Fan and D. R. Kearns (1990) A fluorescence study of the binding of Hoechst 33258 and DAPI to halogenated DNAs. Photochem. Photobiol. 51, 77-86.