DTAF C23H12Cl2N4O5*HCl ~ 492 ~ 514 532 Proteins Prodan C15H17NO ~ 361 ~ 498 227 GFP Varies Varies 26900 Proteins “238 amino acids” Table 2. Examples of fluorophores often used for FRAP experiments. FITC = FluoresceinIsothiocyanate; NBD = nitrobenzoxadiazole; DTAF = fluorescein dichlorotriazine. Table 3. Basic properties of photoactivatable, photoconvertible and photoswitchable fluorescent proteins. Selected FRAP models Empirical Stationary Variance Uniform 3-D et Pixelwise Rectangle 1991) Fourier 2008) Hankel al., multi-photon 2008) transform transform disk FRAP Gaussian (Kapitza FRAP laser (Braeckmans beam (Kubitscheck (Deschout FRAP FRAP FRAP et profile al., (Axelrod (Mazza (Jönsson (Tsay 1985) FRAP et et al., et al., & et al., et 2003 Jacobson, al., (Jonasson 2010) et al., 1994) al., 1976 2008) Additional assumptions 2-D diffusion x x x x x x x Circular bleach region or x x x x x circular symmetry Large bleach area compared x x to bleaching and imaging PSF
DTAF C23H12Cl2N4O5*HCl ~ 492 ~ 514 532 Proteins Prodan C15H17NO ~ 361 ~ 498 227 GFP “238 amino acids” Varies Varies 26900 Proteins Table 2. Examples of fluorophores often used for FRAP experiments. FITC = FluoresceinIsothiocyanate; NBD = nitrobenzoxadiazole; DTAF = fluorescein dichlorotriazine. Table 3. Basic properties of photoactivatable, photoconvertible and photoswitchable fluorescent proteins. Selected FRAP models Empirical (Kapitza et al., 1985) Stationary laser beam (Axelrod et al., 1976 Variance FRAP (Kubitscheck et al., 1994) Uniform disk (Braeckmans et al., 2003 3-D multi-photon FRAP (Mazza et al., 2008) Pixelwise Gaussian profile FRAP (Jonasson et al., 2008) Rectangle FRAP (Deschout et al., 2010) Fourier transform FRAP (Tsay & Jacobson, 1991) Hankel transform FRAP (Jönsson et al., 2008) Additional assumptions 2-D diffusion x x x x x x x Circular bleach region or circular symmetry x x x x x Large bleach area compared to bleaching and imaging PSF x x