By James W Putney
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Extra info for Calcium signaling
Originally estimated at 40-fold , but this was less than it should have been. Subsequent improvement in purification procedures gave an indicator with a 100fold increase in brightness due to calcium binding . This large dynamic range is important for detecting small localized changes in calcium . Fluo-4 is identical in its structure to fluo-3 except for the halogen substitutions on the fluorescein ring . 5 . The lower pKa ensured that the phenolic OH was always ionized within the physiological pH range.
When observed with fura-2, these distinctions tend to be blurred due to the tendency James W. 2005 9:02pm 21 Fluorescent Calcium Indicators Based on BAPTA of high-affinity indicators to saturate or buffer calcium transients. The recent introduction of low-affinity indicators has led to several studies showing large differences in amplitude of calcium transients evoked by different stimuli. This may lead to renewed interest in ‘‘amplitude modulation’’ as an important modality in calcium signaling .
In addition to its brighter fluorescence, fluo-4 is better suited for use with confocal microscopy. With an excitation maxima at 491 nm, fluo-4 is excited much more efficiently than fluo-3 by the 488 nm argon laser line. In addition fluo-4 has greater photostability and better loading characteristics than fluo-3. Despite its improved photostability, fluo-4 has reportedly induced spurious calcium oscillations and caused increased cell death possibly due to the production of James W. 2005 9:02pm 13 reactive oxygen compounds .