Inscrição na biblioteca: Guest
Portal Digital Begell Biblioteca digital da Begell eBooks Diários Referências e Anais Coleções de pesquisa
Journal of Environmental Pathology, Toxicology and Oncology
Fator do impacto: 1.241 FI de cinco anos: 1.349 SJR: 0.356 SNIP: 0.613 CiteScore™: 1.61

ISSN Imprimir: 0731-8898
ISSN On-line: 2162-6537

Journal of Environmental Pathology, Toxicology and Oncology

DOI: 10.1615/JEnvironPatholToxicolOncol.v26.i2.30
pages 83-88

Cell-Substrate Topology upon ALA-PDT Using Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)

Henri-Pierre Lassalle
Hochschule Aalen, Institut für Angewandte Forschung, Anton-Huber-Str. 21, 73430 Aalen, Germany. Permanent address: CRAN UMR 7039, Nancy University, CNRS, CAV, Avenue de Bourgogne, 54511 Vandoeuvre les Nancy, France
Harald Baumann
Hochschule Aalen, Institut für Angewandte Forschung, Anton-Huber-Str. 21, 73430 Aalen, Germany
Wolfgang S. L. Strauss
Institut für Lasertechnologien in der Medizin und Messtechnik, Helmholtzstr. 12, 89081 Ulm, Germany
Herbert Schneckenburger
Hochschule Aalen, Institut für Angewandte Forschung, Anton-Huber-Str. 21, 73430 Aalen, Germany; and Institut für Lasertechnologien in der Medizin und Messtechnik, Helmholtzstr. 12, 89081 Ulm, Germany

RESUMO

Because of the low penetration depth of an evanescent electromagnetic field, total internal reflection fluorescence microscopy (TIRFM) proved to be a powerful technique to examine fluorescent dyes or photosensitizers in close vicinity to the plasma membrane of living cells. In addition, on variation of the angle of incidence of exciting laser light, the penetration depth is varied, so that cell-substrate topology can be examined with nanometer resolution. Using a specific illumination device for TIRFM and a highly sensitive electron multiplying (EM) CCD camera, fluorescence of the photosensitizer protoporphyrin IX (PPIX) was studied in human cancer cells after application of 5-aminolevulinic acid (5-ALA) prior to and after irradiation with sublethal light doses (635 nm, 4 J/cm2). For cells growing on microscope cover slides, cell-substrate distances varied between approximately 20 and 250 nm with a mean distance of approximately 120 nm. On light exposure, these distances generally decreased, and a mean value below 100 nm was attained. Moreover, focal contacts visualized with a fusion protein of yellow fluorescent protein and focal adhesion kinase were maintained on light exposure, i.e., light-induced detachment of cells from their substrate was not likely to occur.


Articles with similar content:

Utility of the F98 Rat Glioma Model for Photodynamic Therapy
Journal of Environmental Pathology, Toxicology and Oncology, Vol.26, 2007, issue 2
Henry Hirschberg, Steen J. Madsen, Khishigzaya Kharkhuu
The Wonders of Silk Fibroin Biomaterials in the Treatment of Breast Cancer
Critical Reviews™ in Eukaryotic Gene Expression, Vol.28, 2018, issue 2
Terin Adali, Pinar Tulay, Nanyak Galam
Total Internal Reflection Microscopy and Atomic Force Microscopy (TIRFM-AFM) to Study Stress Transduction Mechanisms in Endothelial Cells
Critical Reviews™ in Biomedical Engineering, Vol.28, 2000, issue 1&2
Anshu Bagga Mathur, George A. Truskey, W. Monty Reichert
Preface: Special Issue of Photodynamic Therapy and Photodetection with Porphyrin Precursors for the Journal of Environmental Pathology, Toxicology, and Oncology
Journal of Environmental Pathology, Toxicology and Oncology, Vol.26, 2007, issue 2
Alcira Batlle, Qian Peng
Inositol Hexaphosphate Induces Apoptosis by Coordinative Modulation of P53, Bcl-2 and Sequential Activation of Caspases in 7,12 Dimeth-ylbenz [a] anthracene Exposed Mouse Epidermis
Journal of Environmental Pathology, Toxicology and Oncology, Vol.27, 2008, issue 3
Jaya Singh, Krishna P. Gupta