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Synlett 2006(12): 1863-1866  
DOI: 10.1055/s-2006-948165
LETTER
© Georg Thieme Verlag Stuttgart · New York

A Water-Soluble Diaminostilbene Derivative as a Two-Photon Fluorescent Probe

Sheng Yao, Katherine J. Schafer-Hales, Ion Cohanoschi, Florencio E. Hernández, Kevin D. Belfield*
Department of Chemistry and College of Optics and Photonics: CREOL & FPC, University of Central Florida, P.O. Box 162366, Orlando, FL 32816-2366, USA
Fax: +1(407)8232252; e-Mail: kbelfiel@mail.ucf.edu;
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Publication History

Received 23 March 2006
Publication Date:
24 July 2006 (online)

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Abstract

A hydrophilic trans-4,4′-diaminostilbene derivative was prepared by the introduction of amideamino peripheral substituents. Very high aqueous solubility was achieved (>15 mM), making it possible as a fluorescent probe for biological imaging. The relatively straightforward synthetic methodology has great potential to transform even large hydrophobic two-photon absorbing chromophores into hydrophilic derivatives. The fluorescence quantum yield of 0.27 for the diaminostilbene in water is sufficiently high for use as a fluorescent probe. In addition, two-photon fluorescence ­images of NT2 cells stained by this probe were also obtained.

Key words

two-photon - fluorescent probe - PAMAM: hydrophilic - Z-scan - two-photon microscopy

    References and Notes

  • For reviews, see:
  • 1a Rubart M. Circ. Res.  2004,  95:  1154 
    CrossrefGoogle Scholar
  • 1b Heikal AA. Webb WW. Proc. SPIE - Int. Soc. Opt. Eng.  2002,  4812:  1 ; Nonlinear Spectroscopy
    CrossrefGoogle Scholar
  • 1c So PTC. Dong CY. Masters BR. Berland KM. Annu. Rev. Biomed. Eng.  2000,  2:  399 
    CrossrefGoogle Scholar
  • 1d Potter SM. Curr. Biol.  1996,  6:  1595 
    CrossrefGoogle Scholar
  • 2a Xu C. Zipfel W. Shear JB. Williams RM. Webb WW. Proc. Natl. Acad. Sci. U.S.A.  1996,  93:  10763 
    CrossrefGoogle Scholar
  • 2b Xu C. In Confocal and Two-Photon Microscopy   Diaspro A. Wiley-Liss; New York: 2002.  p.75-99  
    CrossrefGoogle Scholar
  • 3a Ohulchanskyy TY. Pudavar HE. Yarmoluk SM. Yashchuk VM. Bergey EJ. Prasad PN. Photochem. Photobiol.  2003,  77:  138 
    CrossrefGoogle Scholar
  • 3b Meltola NJ. Wahlroos R. Soini AE. J. Fluoresc.  2004,  14:  635 
    CrossrefGoogle Scholar
  • 3c Meltola NJ. Soini AE. Hanninen PE. J. Fluoresc.  2004,  14:  129 
    CrossrefGoogle Scholar
  • For example, see:
  • 4a Belfield KD. Morales AR. Kang B.-S. Hales JM. Hagan DJ. Van Stryland EW. Chapela VM. Percino J. Chem. Mater.  2004,  16:  4634 
    CrossrefGoogle Scholar
  • 4b Belfield KD. Morales AR. Hales JM. Hagan DJ. Van Stryland EW. Chapela VM. Percino J. Chem. Mater.  2004,  16:  2267 
    CrossrefGoogle Scholar
  • 4c Rumi M. Ehrlich JE. Heikal AA. Perry JW. Barlow S. Hu Z. McCord-Maughon D. Parker TC. Rockel H. Thayumanavan S. Marder SR. Beljonne D. Bredas J.-L. J. Am. Chem. Soc.  2000,  122:  9500 
    CrossrefGoogle Scholar
  • 4d Albota M. Beljonne D. Bredas J.-L. Ehrlich JE. Fu J.-Y. Heikal AA. Hess SE. Kogej T. Levin MD. Marder SR. McCord-Maughon D. Perry JW. Rockel H. Rumi M. Subramaniam G. Webb WW. Wu X.-L. Xu C. Science  1998,  281:  1653 
    CrossrefGoogle Scholar
  • 4e Reinhardt BA. Brott LL. Clarson SJ. Dillard AG. Bhatt JC. Kannan R. Yuan L. He GS. Prasad PN. Chem. Mater.  1998,  10:  1863 
    CrossrefGoogle Scholar
  • 4f Abbotto A. Beverina L. Bozio R. Facchetti A. Ferrante C. Pagani GA. Pedron D. Signorini R. Org. Lett.  2002,  4:  1495 
    CrossrefGoogle Scholar
  • For reviews see:
  • 5a Esfand R. Tomalia DA. Drug Discov. Today  2001,  6:  427 
    CrossrefPubMedGoogle Scholar
  • 5b Boas U. Heegaard PMH. Chem. Soc. Rev.  2004,  33:  43 
    CrossrefPubMedGoogle Scholar
  • For examples, see:
  • 6a Day PN. Nguyen KA. Pachter R. J. Phys. Chem. B  2005,  109:  1803 
    CrossrefGoogle Scholar
  • 6b Drobizhev M. Karotki A. Dzenis Y. Rebane A. Suo Z. Spangler CW. J. Phys. Chem. B  2003,  107:  7540 
    CrossrefGoogle Scholar
  • 6c Antonov L. Kamada K. Ohta K. Kamounah FS. Phys. Chem. Chem. Phys.  2003,  5:  1193 
    CrossrefGoogle Scholar
  • 6d Das GP. Yeates AT. Dudis DS. Chem. Phys. Lett.  2002,  361:  71 
    CrossrefGoogle Scholar
  • 6e Wang X. Wang D. Zhou GY. Yu W. Zhou Y. Fang Q. Jiang M. J. Mater. Chem.  2001,  11:  1600 
    CrossrefGoogle Scholar
  • 7 Prepared according to a literature procedure, see: Calvin M. Buckles RE. J. Am. Chem. Soc.  1948,  62:  3324 
    Google Scholar
  • 8a Kim Y.-W. Jin J.-L. Jin MY. Choi K.-Y. Kim J.-J. Zyung T. Polymer  1997,  38:  2269 
    CrossrefGoogle Scholar
  • 8b Tsubokawa N. Satoh T. Murota M. Sato S. Shimizu H. Polym. Adv. Technol.  2001,  12:  596 
    CrossrefGoogle Scholar
  • 10 Zhou G. Wang X. Wang D. Shao Z. Jiang M. Appl. Opt.  2002,  41:  1120 
    Google Scholar
  • 11a The 2PA cross-sections measurement followed the procedure described in: Hernandez FE. Belfield KD. Cohanoschi I. Chem. Phys. Lett.  2004,  391:  22 
    CrossrefGoogle Scholar
  • 11b

    interestingly, two-photon cross-section of compound 1 is 3500 GM measured by picosecond laser sources, about one order of magnitude higher than that measured by femtosecond laser source. Similar phenomena have been reported in results from nanosecond laser sources by Prasad and co-workers; for examples, see:
    Crossref
  • 11c Chung S. Kim K. Lin T. He GS. Swiatkiewicz J. Prasad PN. J. Phys. Chem.  1999,  103:  10741 
    CrossrefGoogle Scholar
  • 11d Kim OK. Lee K.-S. Woo HY. Kim K.-S. He GS. Swiatkiewicz J. Prasad PN. Chem. Mater.  2000,  12:  284 
    CrossrefGoogle Scholar
  • 11e He GS. Lin T.-C. Prasad PN. Kannan R. Vaia RA. Tan L.-S. J. Phys. Chem. B  2002,  106:  11081 
    CrossrefGoogle Scholar
  • 11f

    More detailed experimental investigation is underway to understand the mechanism.
    Crossref
  • 12 The sample preparation and multiphoton fluorescence imaging followed the procedure described in: Schafer-Hales KJ. Belfield KD. Yao S. Frederiksen PK. Hales JM. Kolattukudy PE. J. Biomed. Opt.  2005,  10:  051402 
    CrossrefGoogle Scholar
9

Experimental Procedure.
1H NMR spectra were recorded on a Varian Mercury-300 NMR (300 MHz) spectrometer using TMS as the internal standard. Chemical shifts (δ) are reported in parts per million (ppm). Elemental analyses were performed by Atlantic Microlab, Inc. Norcross, GA. UV/visible spectra were recorded on an Agilent 8453 spectrophotometer using standard 1 cm path length cuvettes. Steady-state and fluorescence excitation anisotropy measurements were performed on a PTI Quantamaster spectrofluorometer. Fluorescence quantum yields were determined relative to 9,10-diphenylanthracone in cyclohexane.
Preparation of 3,3′,3′′,3′′′-[( E )-Ethene-1,2-diylbis(4,1-phenylenenitrilo)]tetrapropionic Acid Tetraethyl Ester ( 4). A mixture of 4,4′-diaminostilbene (6.3 g, 0.03 mol), freshly distilled ethyl acrylate (30 mL, 0.27 mol), and AcOH (4.5 mL) was refluxed for 48 h. The excess ethyl acrylate was removed under reduced pressure. The residue was purified by column chromatography using hexane-EtOAc (2:1) as eluent. Solvent removal and recrystallization from 95% EtOH afforded 7.2 g of pure product (39% yield). 1H NMR (300 MHz, CDCl3): δ = 7.35 (d, J = 7.8 Hz, 4 H, PhH), 6.82 (s, 2 H, CH=CH), 6.67 (d, J = 7.8 Hz, 4 H, PhH), 4.13 (q, J = 6.9 Hz, 8 H, OCH2), 3.67 (t, J = 7.1 Hz, 8 H, NCH2), 2.59 (t, J = 6.8 Hz, 8 H, C=OCH2), 1.26 (t, J = 7.2 Hz, 12 H, CH3). Anal. Calcd for C34H46N2O8 (610.74): C, 66.86; H, 7.59; N, 4.59. Found: C, 67.02; H, 7.62; N, 4.65.
Preparation of 3,3′,3′′,3′′′-[( E )-Ethene-1,2-diylbis(4,1-phenylenenitrilo)]tetrakis[ N -(2-aminoethyl)]propan-amide ( 1).
Diaminoethylene (6.67 g, 111 mmol) was added to a solution of 4 (0.61 g, 1.0 mmol) in MeOH (3.2 mL), and the mixture was reacted at r.t. under N2 for 60 h. The solvent and the remaining diaminoethylene were removed by distillation at 40 °C under reduced pressure. The yellow solid obtained was washed with Et2O and collected by suction filtration to afford 0.63 g of product (94% yield). NMR analysis indicated this product is a mixture of trans- and cis-isomers, which was recrystallized in EtOH-hexane for three times to give a pure trans-product; mp 180-181 °C. 1H NMR (300 MHz, D2O): δ = 7.25 (d, J = 5.1 Hz, 4 H, PhH), 6.76 (d, J = 3.9 Hz, 2 H, CH=CH), 6.64 (d, J = 5.4 Hz, 4 H, PhH), 3.46 (t, J = 3.5 Hz, 8 H, CH2), 2.95 (t, J = 3.6 Hz, 8 H, CH2), 2.40 (t, J = 3.6 Hz, 8 H, CH2), 2.29 (t, J = 3.8 Hz, 8 H, CH2). Anal. Calcd for C34H54N10O4·3H2O (720.90): C, 56.65; H, 8.40; N, 19.43. Found: C, 56.82; H, 8.25; N, 19.07.