[EN] TWO-PHOTON PROBE FOR REAL-TIME MONITORING OF INTRACELLULAR CALCIUM IONS, METHOD FOR PREPARING THE PROBE AND METHOD FOR REAL-TIME MONITORING OF INTRACELLULAR CALCIUM IONS USING THE PROBE<br/>[FR] SONDE À DEUX PHOTONS POUR SURVEILLANCE EN TEMPS RÉEL D'IONS DE CALCIUM INTRA-CELLULAIRE, PROCÉDÉ DE PRÉPARATION DE LA SONDE ET PROCÉDÉ DE SURVEILLANCE EN TEMPS RÉEL DES IONS DE CALCIUM INTRA-CELLULAIRE UTILISANT LA SONDE
申请人:UNIV IND & ACAD COLLABORATION
公开号:WO2009031734A1
公开(公告)日:2009-03-12
A two-photon probe for real-time monitoring of intracellular calcium ions is provided. The two-photon probe is very suitable for real-time imaging of intracellular calcium ions, shows 20~5 O-fold TPEF enhancement in response to Ca2+, has a dissociation constant (Kdτp) of 0.14 ± 0.02 to 0.25 ± 0.03 μM, and emits 5-fold stronger TPEF than currently available one-photon fluorescent Ca2+ probes. Unlike the previously available probes, the two-photon probe can selectively detect dynamic levels of intracellular free Ca2+ in live cells and living tissues without interference from other metal ions and from the membrane-bound probes. Moreover, the two- photon probe is capable of monitoring the calcium waves at a depth of 100-300 μm in live tissues for 1,100-4,000 s using two-photon microscopy (TPM) with no artifacts of photo-bleaching. Further provided are a method for preparing the two-photon probe and a method for real-time monitoring of intracellular calcium ions using the two-photon probe.
提供了一种用于实时监测细胞内钙离子的双光子探针。这种双光子探针非常适合实时成像细胞内钙离子,对Ca2+的响应显示20~50倍的TPEF增强,其解离常数(Kdτp)为0.14 ± 0.02至0.25 ± 0.03μM,并且发出比当前可用的单光子荧光Ca2+探针强5倍的TPEF。与先前可用的探针不同,双光子探针可以选择性地检测活细胞和活组织中的细胞内游离Ca2+的动态水平,而不受其他金属离子和膜结合探针的干扰。此外,双光子探针能够使用双光子显微镜(TPM)在活组织中以100-300μm深度监测钙波长达1,100-4,000秒,而不会出现光漂白的伪影。还提供了制备双光子探针的方法以及使用双光子探针进行实时监测细胞内钙离子的方法。