A near-infrared fluorescent calcium probe: a new tool for intracellular multicolour Ca2+ imaging

Matsui, Akihiro; Umezawa, Keitaro; Shindo, Yutaka; Fujii, Tomohiko; Citterio, Daniel; Oka, Kotaro; Suzuki, Kôji

doi:10.1039/c1cc14045k

Cited by 81 publications

(40 citation statements)

References 17 publications

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“…The red or far-red wavelength region is especially attractive for fluorescence imaging, affording higher tissue penetration due to reduced scattering, low absorption by endogenous biomolecules, and lower phototoxicity, and also allows monitoring of Ca 2+ in cells or tissues expressing yellow-or green-colored fluorescent proteins or labeled with other green-colored fluorophores [20][21][22]. Several new red-to-NIR fluorescent probes for cytoplasmic Ca 2+ , such as Quest Rhod-4 AM, CaSiR-1 [23], KFCA [24], and Calcium Rubies [22,25] have recently been reported [20]. We developed a red (ca.…”

Section: Introductionmentioning

confidence: 99%

Development of practical red fluorescent probe for cytoplasmic calcium ions with greatly improved cell-membrane permeability

et al. 2016

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Fluorescence imaging of calcium ions (Ca(2+)) has become an essential technique for investigation of signaling pathways involving Ca(2+) as a second messenger. But, Ca(2+) signaling is involved in many biological phenomena, and therefore simultaneous visualization of Ca(2+) and other biomolecules (multicolor imaging) would be particularly informative. For this purpose, we set out to develop a fluorescent probe for Ca(2+) that would operate in a different color region (red) from that of probes for other molecules, many of which show green fluorescence, as exemplified by green fluorescent protein (GFP). We previously developed a red fluorescent probe for monitoring cytoplasmic Ca(2+) concentration, based on our established red fluorophore, TokyoMagenta (TM), but there remained room for improvement, especially as regards efficiency of introduction into cells. We considered that this issue was probably mainly due to limited water solubility of the probe. So, we designed and synthesized a red-fluorescent probe with improved water solubility. We confirmed that this Ca(2+) red-fluorescent probe showed high cell-membrane permeability with bright fluorescence. It was successfully applied to fluorescence imaging of not only live cells, but also brain slices, and should be practically useful for multicolor imaging studies of biological mechanisms.

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Section: Introductionmentioning

confidence: 99%

Development of practical red fluorescent probe for cytoplasmic calcium ions with greatly improved cell-membrane permeability

et al. 2016

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“…Attempts to solve this problem by the simultaneous imaging of fluo-3 and fura-red [24] have enjoyed limited success. This situation has prompted the development of new probes with improved characteristics such as ICPBC [25], calcium ruby [26, 27], CaSiR1 [28] and KFCA [29]. As they have become available only recently, very few examples of their applications have been described.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and properties of Asante Calcium Red—A novel family of long excitation wavelength calcium indicators

Hyrc

Minta²,

Escamilla³

et al. 2013

Cell Calcium

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Although many synthetic calcium indicators are available, a search for compounds with improved characteristics continues. Here, we describe the synthesis and properties of Asante Calcium Red-1 (ACR-1) and its low affinity derivative (ACR-1-LA) created by linking BAPTA to seminaphthofluorescein. The indicators combine a visible light (450–540 nm) excitation with deep-red fluorescence (640 nm). Upon Ca2+ binding, the indicators raise their fluorescence with longer excitation wavelengths producing higher responses. Although the changes occur without any spectral shifts, it is possible to ratio Ca2+-dependent (640 nm) and quasi-independent (530 nm) emission when using visible (<490 nm) or multiphoton (~780 nm) excitation. Therefore, both probes can be used as single wavelength or, less dynamic, ratiometric indicators. Long indicator emission might allow easy [Ca2+]i measurement in GFP expressing cells. The indicators bind Ca2+ with either high (Kd=0.49±0.07 μM; ACR-1) or low affinity (Kd=6.65±0.13 μM; ACR-1-LA). Chelating Zn2+ (Kd =0.38±0.02 nM) or Mg2+ (Kd ~5 mM) slightly raises and binding Co2+ quenches dye fluorescence. New indicators are somewhat pH-sensitive (pKa=6.31±0.07), but fairly resistant to bleaching. The probes are rather dim, which combined with low AM ester loading efficiency, might complicate in situ imaging. Despite potential drawbacks, ACR-1 and ACR-1-LA are promising new calcium indicators.

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“…Suzuki et al. reported a BODIPY based NIR probe 12 for the intracellular Ca 2+ ions (Figure ) . In fluorescence, 12 did not exhibit any emission, however, upon the addition of Ca 2+ ions, it displayed 120‐fold fluorescence enhancement at 670 nm ( Φ =0.24).…”

Section: Carboxylate Groups Appended Probesmentioning

confidence: 99%

Design and Applications of Small Molecular Probes for Calcium Detection

Roopa

Kumar

2019

Chemistry An Asian Journal

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The physiological significance of calcium ions such as the role in cellular signalling, cell growth,e tc. have driven the development of methods to detect and monitort he level of Ca 2 + ions, both in vivo and in vitro. Although various approaches for the detection of calcium ions have been reported, methods based on small molecular fluorescent probesh ave uniquea dvantages including smallp robe size, easy monitoring of detection processes and applicability in biological systems. In this review article, we will discusst he progress in the development of Ca 2 + -binding fluorescent probesb yt aking into accountt he types of chelating groups that have been employed for Ca 2 + binding.Scheme1.Designs trategyo faf luorescentsensor.[a] Dr.Roopa

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A near-infrared fluorescent calcium probe: a new tool for intracellular multicolour Ca2+ imaging

Cited by 81 publications

References 17 publications

Development of practical red fluorescent probe for cytoplasmic calcium ions with greatly improved cell-membrane permeability

Development of practical red fluorescent probe for cytoplasmic calcium ions with greatly improved cell-membrane permeability

Synthesis and properties of Asante Calcium Red—A novel family of long excitation wavelength calcium indicators

Design and Applications of Small Molecular Probes for Calcium Detection

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