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Although excellent research tools, the ultrafast laser systems are poorly suited for routine laboratory instrumentation, simply due to their high price. Our research group has shown tat application of two-photon excitation need not to be limited to those laboratories that can afford the price and maintenance of ultrafast laser systems. With certain compromises and well-designed experiments, low-cost lasers can be used in utilizing the positive properties of two-photon excitation. The purpose of this publication is to show examples of bioanalytical applications that are possible using low-cost lasers. A bioaffinity assay using microparticles as bioactive carriers and assay for characterization of cell surface antigens are presented. By direct comparison of the example assays in two instrumental set-ups, a quantity is given to the 'compromises' that are made by replacing an ultrafast system with low cost equipment.
Słowa kluczowe
Wydawca
Czasopismo
Rocznik
Tom
Strony
39--44
Opis fizyczny
Bibliogr. 15 poz.
Twórcy
autor
- Laboratory of Biophysics, Institute of Biomedicine, University of Turku Po Box 123, FIN-20521 Turku, Finland
autor
- Laboratory of Biophysics, Institute of Biomedicine, University of Turku Po Box 123, FIN-20521 Turku, Finland
autor
- Laboratory of Biophysics, Institute of Biomedicine, University of Turku Po Box 123, FIN-20521 Turku, Finland
autor
- Laboratory of Biophysics, Institute of Biomedicine, University of Turku Po Box 123, FIN-20521 Turku, Finland
Bibliografia
- 1. W. Denk, J.H. Strickler, and W.W. Webb, “Two-photon-laser scanning fluorescence microscopy” Science 248, 73-76 (1990).
- 2. M. Gëppert-Mayer, "Über Elementarakte mit zwei Quantensprüngen", Ann. Phys. 9, 273 (1931).
- 3. W. Denk, D.W. Piston, and W.W. Webb, "Two-photon molecular excitation in laser scanning microscopy", in Hand-book of Confocal Microscopy, pp. 445-458, edited by J.B. Pawley, Plenum, New York, 1995.
- 4. P.E. Hänninen, E. Soini, and S.W. Heli "Continuous wave excitation two-photon fluorescence microscopy", Journal of Micr. 176, 222-225 (1994).
- 5. P.E. Hänninen, M. Schrader, E. Soini, and S.W. Heli, "Two-photon excitation microscopy using a semiconductor laser", Bioimaging 3, 70-75 (1995).
- 6. E.L. Florin, J.K.H. Horber, and E.H.K. Stelzer, "High-resolution axial and lateral position sensing using two-photon excitation of fluorophores by a continuous-wave Nd alpha YAG laser" Appl. Phys. Lett. 69, 446-448 (1996).
- 7. K. Kenig, "Laser tweezers are sources of two-photon excitation" Cell. & Mol. Biol. 44, 721-733 (1998).
- 8. P. Hänninen, A. Soini, N. Meltola, J. Soini, J. Soukka, and E. Soini, "A new microvolume technique for bioaffinity assay using two-photon excitation", Nature Biotec. 18, 548-550 (2000).
- 9. M.E. Waris, N.J. Meltola, J.T. Soini, E. Soini, O.J. Peltola, and P.E. Hänninen, "Two-photon excitation fluorometric measurement of homogeneous microparticle immunoassay for C-reactive protein", Anal. Biochem. 309, 67-74 (2002).
- 10. A. Schönle, and S.W. Hell, "Heating by absorption in the focus of an objective lens", Opt. Lett. 23, 325-327 (1998).
- 11. K.M. Berland, P.T.C. So, and E. Gratton, "Two-photon fluorescence correlation spectroscopy: method and application to the intracellular environment", Biophys. Journal 68, 694-701 (1995) .
- 12. A. Van Orden, H. Cai, P.M. Goodwin, and R.A. Keller, "Efficient detection of single DNA fragments in flowing sample streams by two-photon fluorescence excitation", Anal. Chem. 71, 2108-2116 (1999).
- 13. E. Soini, P. Hanninen, and J. Soini, "Biospecific multiparametric assay method", FI-101829, EP0804732, DE 69615818.3-08, Japan 8-522062, US 5,891738. priority 03/1995 (1995).
- 14. K.G. Heinze, A. Koltermann, and P. Schwille, "Simultaneous two-photon excitation of distinct labels for dual-colour fluorescence cross-correlation analysis", Proc. Natl. Acad. Sci. 97, 10377-10382 (2000).
- 15. J.T. Soini, J.M. Soukka, E. Soini, and P.E. Hänninen, "Two-photon excitation microfluorometer for multiplexed single-step bioaffinity assays", Rev. Sci. Instr. 73, 2680-2685 (2002).
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Bibliografia
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