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pH Fluorescent Probes: Chlorinated Fluoresceins

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Abstract

A series of regiospecific chlorinated fluoresceins have been synthesized by the reaction of the regiospecific chlorinated resorcinols with chlorinated phthalic anhydride. The regioisomers were successfully separated by chromatography. The photophysical properties of the obtained chlorinated fluoresceins were examined and found their absorption and emission maxima at long wavelength with high fluorescence quantum yield. Especially, pH-dependent properties of chlorinated fluoresceins have been studied in detail. These compounds show strongly pH-sensitive range of 3.5–7.0, and have lower pK a values than fluorescein. Furthermore, their fluorescent intensity could reach the maximum in the physiological environment of pH range 6.8–7.4. Due to higher fluorescence quantum yield and lower pK a values, chlorinated fluoresceins will be expected to be used as excellent pH fluorescent probes for pH measurement of the acidic cell.

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References

  1. Grinstein S, Goetz JD (1985) Control of free cytoplasmic calcium by intracellular pH in rat lymphocytes. Biochim Biophys Acta 819(2):267–270

    Article  PubMed  CAS  Google Scholar 

  2. Vergne I, Constant P, Lan, elle G (1998) Phagosomal pH determination by dual fluorescence flow cytometry. Anal Biochem 255:127–132

    Article  PubMed  CAS  Google Scholar 

  3. Diwu Z, Twu JJ, Yi G, Lavis LD, Chen Y, Cassutt KJ (2003) Fluorescent pH indicators for intracellular assays. US 20030068668A1

  4. Jin SY, Xu ZC, Chen JP, Liang XM, Wu YN, Qian XH (2004) Determination of organophosphate and carbamate pesticides based on enzyme inhibition using a pH-sensitive fluorescence probe. Anal Chim Acta 523(1):117–123

    Article  CAS  Google Scholar 

  5. Galande AK, Weissleder R, Tung CH (2006) Fluorescence probe with a pH-sensitive trigger. Bioconjugate Chem 17(2):255–257

    Article  CAS  Google Scholar 

  6. Haugland RP (2002) Handbook of fluorescent probes and research products, 9th ed. Molecular Probes, Eugene, pp 830–833

  7. Song L, Hennink EJ, Young T, Tanke HJ (1995) Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy. Biophys J 68(6):2588–2600

    PubMed  CAS  Google Scholar 

  8. Song L, Varma CAG, Verhoeven JW, Tanke HJ (1996) Influence of the triplet excited state on the photobleaching kinetics of fluorescein in microscopy. Biophys J 70(6):2959–2968

    Article  PubMed  CAS  Google Scholar 

  9. Guilbault GG (ed) (1990) In: Practical fluorescence, chapter1. Marcel Dekker, NewYork

  10. Graber ML, Dilillo DC, Friedman BL, Enrique PM (1986) Characteristics of fluoroprobes for measuring intracellular pH. Anal Bio 156:202–212

    Article  CAS  Google Scholar 

  11. Millar D, Uttamal M, Henderson R, Keeper A (1998) Electrochemical immobilization of a pH sensitive fluorescein derivative: synthesis and characterization of a fluorescein-derivatised polythiophene. Chem Commun 447–478

  12. Whitaker JE, Haugland RP, Prendergast FG (1991) Spectral and photophysical studies of benzo[c]xanthene dyes: dual emission pH sensors. Anal Bio 194:3330–340

    Google Scholar 

  13. Lyttle MH, Carter TG, Cook RM (2001) Improved synthetic procedures for 4,7,2′,7′-tetrachloro- and 4′,5′-dichloro-2′,7′-dimethoxy-5 (and 6)-carboxy- fluoresceins. Org Pro Res Dev 5:45–49

    Article  CAS  Google Scholar 

  14. Demas JN, Crosby GA (1971) Measurement of photoluminescence quantum yields. J Phy Chem 75:991–1024

    Article  Google Scholar 

  15. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    PubMed  CAS  Google Scholar 

  16. Babcock DF (1983) Examination of the intracellular ionic environment and of ionophore action by null point measurements employing the fluorescein chromophore. J Biol Chem 258:6380–6389

    PubMed  CAS  Google Scholar 

  17. Ge FY, Yan XL, Yan FY, Pan HY, Chen LG (2005) New fluorescent labels: 4-and 7-chlorofluorescein. J Fluorescence 15(6):829–833

    Article  CAS  Google Scholar 

  18. Guilbault GG (ed) (1990) In: Practical fluorescence, chapter 6. Marcel Dekker, NewYork

  19. Fujio M, Mciver RT, Taft RW (1981) Effects on the acidities of phenols from specific substituent–solvent interactions. Inherent substituent parameters from gas-phase acidities.. J Am Chem Soc 103:4017–4029

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, Donghua University, Shanghai, 201620, People’s Republic of China

    Feng-Yan Ge

  2. Key Laboratory of Eco-textile (Donghua University), Ministry of Education, Shanghai, 201620, People’s Republic of China

    Feng-Yan Ge

  3. School of Chemical Engineering and technology, Tianjin University, Tianjin, 300072, People’s Republic of China

    Li-Gong Chen

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  1. Feng-Yan Ge
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  2. Li-Gong Chen
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Correspondence to Feng-Yan Ge.

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Ge, FY., Chen, LG. pH Fluorescent Probes: Chlorinated Fluoresceins. J Fluoresc 18, 741–747 (2008). https://doi.org/10.1007/s10895-007-0305-y

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  • DOI: https://doi.org/10.1007/s10895-007-0305-y

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