The emission maximum of the fluorescent specstrum shifts from 485 nm to 410 nm with Ca complexation, while the excitation maximum remains at around 300 nm. Thus two separate wavelengths in the emission specstrum, usually at 480 nm and 410 nm, are measured in ratiometry of Indo 1 experiment, making Indo 1 suitable for the ratiometry using flow cytometor etc. Indo 1 is reported to have a better property in the compartmentation after loaded into cells than Fura 2 .
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References G. Grynkiewicz, M. Poenie, R. Y. Tsien, A new generation of calcium indicators with greatly improved fluorescence properties, J. Biol. E. Niggli, D. W. Piston, M. S. Kirby, H. Cheng, D. R. Sandison, W. W. Webb, W. J. Lederer, A confocal laser scanning microscope designed for indicators with ultraviolet excitation wavelengths, Am. L. Darjania, N. Curvetto, S. Delmastro, Loading of Vicia faba guard cell protoplasts with Indo 1 to measure cytosolic calcium concentration, Plant Physiol. R. Brandes, V. M. Figueredo, S. A. Camacho, M. W. Weiner, Compensation for changes in tissue light absorption in fluorometry of hypoxic perfused rat hearts, Am. J. Physiol .