Lasers in Medical Science

, Volume 25, Issue 4, pp 545–550 | Cite as

Laser-induced effects in different biological samples

Original Article

Abstract

Experiments were carried out on cancerous HeLa cells and blood serum using a double integrating sphere and a He-Ne laser to investigate the optical properties and cellular effects due to photodynamic therapy (PDT). In the first experiment, HeLa cells were exposed to Photofrin at concentrations of 0, 10, 20, 30, 50 and 112.4 µg/ml at an irradiance of 0.2 W/cm2 using diode laser light. Using a confocal microscope, cell debris and morphological changes in HeLa cells were recorded at different Photofrin concentrations. The results showed cell debris in HeLa cells at the highest concentration of Photofrin. In a second experiment, photobleaching was observed in HeLa cells in the presence of various concentrations of 5-aminolaevulinic acid ranging from 0–50 µg/ml. There was progressive degradation of the 635 nm peak during continuous laser irradiation at an irradiance of 0.2 W/cm2. We conclude that cells demonstrating high initial fluorescence undergo bleaching at a faster rate than those with lower fluorescence. Finally in a third experiment, cancerous and noncancerous blood serum was irradiated at an irradiance of 0.1 W/cm2 using a He-Ne laser in conjunction with a double integrating sphere system. Forward and back scattering of normal and malignant serum showed an exponential decrease in fluorescence amplitude. The results indicate that there is notable amplitude difference between malignant and normal blood serum with malignant blood serum showing decreased scattering. These results have important implications for photodiagnosis and photodynamic therapy.

Keywords

Photodynamic therapy HeLa cells Photobleaching effect Confocal microscopy Cell debris 

References

  1. 1.
    Petrova GP, Petrusevich YuM, Boiko AV, Ivanov AV, Papish EA, Khlapov VP, Fedorova KV (2007) Changing optical properties of blood serum proteins in case of oncological disease. In: Dumitras DC, Dinescu M, Konov VI (eds) Advanced Laser Technologies 2006. Proceedings of SPIE, vol. 6606, pp 66061J–66061JGoogle Scholar
  2. 2.
    Roggan A, Minet O, Schroder C, Muller O (1994) Determination of optical tissue properties with double integrating sphere technique and Monte Carlo simulations. In: Tuchin VV (ed) Cell and Biotissue Optics: Applications in Laser Diagnostics and Therapy. Proceedings of SPIE, vol. 2100, pp 42–56Google Scholar
  3. 3.
    Yavari N, Dam JS, Antonsson J, Wårdell K, Andersson-Engels S (2005) Measurements of optical properties of pig brain tissue in vitro using a novel compact device. In: Depeursinge CD (ed) Novel Optical Instrumentation for Biomedical Applications II. Proceedings of SPIE, vol. 5864, p 58640HGoogle Scholar
  4. 4.
    Cheong WF, Prahl SA, Welch AJ (1990) A review of the optical properties of biological tissues. IEEE J Quantum Electron 26:2166–2185CrossRefGoogle Scholar
  5. 5.
    Groenhuis RAJ, Ten Bosch JJ, Ferwerda HA (1983) Scattering and absorption of turbid materials determined from reflection measurements. Appl Opt 22:2463–2467CrossRefPubMedGoogle Scholar
  6. 6.
    Patterson MS, Chance B, Wilson BC (1989) Time resolved reflectance and transmittance for the noninvasive measurement of tissue optical properties. Appl Opt 28:2331–2336CrossRefGoogle Scholar
  7. 7.
    Gallegos ER, Rodríguez ID, Guzmán LAM, Zapata AJP (1999) In vitro study of biosynthesis of protoporphyrin ix induced by δ-aminolevulinic acid in normal and cancerous cells of the human cervix. Arch Med Res 30:163–170CrossRefPubMedGoogle Scholar
  8. 8.
    Karu T, Pyatibrat L, Kalendo G (1995) Irradiation with He-Ne laser increases ATP level in cells cultivated in vitro. J Photochem Photobiol B Biol 27:219–223CrossRefGoogle Scholar
  9. 9.
    Castropazos MD, Soares CP, Silva NSD et al (2003) Ultrastructural effects of two phthalocyanines in CHO-K1 and HeLa cells after laser irradiation. Biocell 27:301–309Google Scholar
  10. 10.
    Atif M, Firdous S, Khurshid A, Noreen L, Zaidi SSZ, Ikram M (2009) In vitro study of 5 Aminolevulinic acid (5-ALA) based photodynamic therapy for apoptosis in human cervical HeLa cell line. Laser Physics Lett 6(12):886–891CrossRefGoogle Scholar
  11. 11.
    Atif M, Dyer PE, Snelling HV, Paget T, Stringer MR (2007) Two-photon excitation studies of mTHPC photosensitizer and photodynamic activity in an epithelial cell line. Photodiagnosis and Photodynamic Therapy Amsterdam: Elsevier 4(2):106–111CrossRefGoogle Scholar
  12. 12.
    Atif M, Stringer MR, Cruse-Sawyer JE, Dyer PE, Brown SB (2005) The influence of intracellular mTHPC concentration upon photobleaching dynamics. Photodiagnosis Photodyn Ther 2(3):235–238CrossRefGoogle Scholar
  13. 13.
    Atif M, Stringer MR, Cruse-Sawyer JE, Brown SB (2004) Fluence-rate effects upon mTHPC photobleaching in a formalin-fixed cell system. Photodiagnosis Photodyn Ther 1(2):173–180Google Scholar
  14. 14.
    Atif M, Stringer MR, Cruse-Sawyer JE, Brown SB (2003) Intracellular fluorescence photobleaching dynamics of mTHPC. Lasers Med Sci 18(1):S51CrossRefGoogle Scholar
  15. 15.
    Schneider M, Graschew G, Roelofs TA, Balanos E, Rakowsky S, Sinn HJ, Schlag PM (2000) Multiphoton excitation and photodynamic activity of macromolecular derivatized mTHPC. In: Dougherty TJ (ed) Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy IX. Proceedings of SPIE, vol. 3909, pp 60–65Google Scholar
  16. 16.
    Black JF, Barton JK (2004) Chemical and structural changes in blood undergoing laser photocoagulation. Photochem Photobiol 80:89–97CrossRefPubMedGoogle Scholar
  17. 17.
    Bagdonas S, Ma LW, Iani A, Rotomskis R, Juzenas P, Moan J (2000) Phototransformations of ALA induced PpIX in vitro: a spectroscopic study. Photochem Photobiol 72:186–192CrossRefPubMedGoogle Scholar
  18. 18.
    Moan J, Streckte G, Bagdonas S, Bech O, Berg K (1997) Photobleaching of PpIX in cells incubated with 5-ALA. Int J Cancer 70:90–97CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag London Ltd 2010

Authors and Affiliations

  1. 1.Biophotonics LaboratoryNational Institute of Lasers and OptronicsIslamabadPakistan

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