Optical imaging of Hsp70 gene expression following thermal laser injury

Josh T. Beckham; J. A. Baran; A. D. Izzo; E. Duco Jansen

doi:10.1117/12.434723

9 July 2001 Optical imaging of Hsp70 gene expression following thermal laser injury

Josh T. Beckham, J. A. Baran, A. D. Izzo, E. Duco Jansen

Proceedings Volume 4257, Laser-Tissue Interaction XII: Photochemical, Photothermal, and Photomechanical; (2001) https://doi.org/10.1117/12.434723
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

In the past the effects of laser irradiation upon tissue have been assessed based on structural and macroscopic characteristics such as temperature, pressure and tissue mass removal. However, the effects of laser irradiation on a cellular level are not well understood and it is postulated that cellular injury caused by laser treatment may affect the efficacy of the laser procedure. In this research we have used an alternative method of detecting injury by targeting the heat shock protein (Hsp70). A stable cell line was generated containing the luciferase reporter gene attached to the heat shock protein (Hsp70). After thermal injury luciferase is produced in tandem with the heat shock protein to emit bioluminescence at 563 nm. The luminescence was quantified with a photon counting ICCD camera system. The heat shock to bring about Hsp70 transcription was created by immersing the cells in a water bath or by irradiating the cells with a Holmium:YAG pulsed laser (λ= 2.1 μm, τ _p = 250 μs). For the laser experiments, radiant exposures varied from 5 to 30 mJ/mm₂ and the number of pulses varied at 15, 25 and 35. The peak expression of luciferase was found to be 3 to 4 hours post heat shock for lower exposures but increase to between 6 and 9 hours if higher radiant energies are used. An experiment was also done to assess to what extent the cellular response to heat followed the Arrhenius rate process.

Citation Download Citation

Josh T. Beckham, J. A. Baran, A. D. Izzo, and E. Duco Jansen "Optical imaging of Hsp70 gene expression following thermal laser injury", Proc. SPIE 4257, Laser-Tissue Interaction XII: Photochemical, Photothermal, and Photomechanical, (9 July 2001); https://doi.org/10.1117/12.434723

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