1,3-alternate coumarin substituted thiacalix［4］arene fluorescent probe 1 was synthesised from 1,3-alternate diethyl thiacalix［4］arene and 7-hydroxycoumarin by-step reactions. In DMSO/H2O (φ, 3/7, pH 7) solution, the strong fluorescence emission and UV absorption of probe 1 can be selectively quenched or significantly enhanced by Fe3+ ion. The probe 1 showed high Fe3+ selective fluorescence quenching or absorption enhancement over commonly coexistent metal ions in neutral aqueous media, and the limit of detections were obtained as low as 10－8 mol·L-1 of Fe3+ by fluorescence and absorption spectrometry. Spectral titration, isothermal titration calorimetry and mass spectrometry were showed that probe and Fe3+ form 1∶1 complexes, the constant up to 105 L·mol-1 and coordinate process was spontaneous by the mole binding free energy and entropy of probe with Fe3+. In addition, the probe can identification bovine hemoglobin (BHb) over other proteins through quenched its fluorescence in DMSO/H2O (φ, 1/9, Tris-HCl, pH 7, 0.1 mol·L-1 NaCl) media. The limit of detection was obtained as 0.12 μg·mL-1 of BHb, as well as a linearity of 0.2～3.0 μg·mL-1, indicating the probe of high sensitivity and quantitation range. It can be used as a selective recognition Fe3+ and BHb of thiacalix<参考文献原文>arene fluorescent probe.