Dynamics of photoinduced cell plasma membrane injury

W. P. Thorpe, M. Toner, R. M. Ezzell, R. G. Tompkins, M. L. Yarmush

Research output: Contribution to journalArticlepeer-review

49 Scopus citations

Abstract

We have developed a video microscopy system designed for real-time measurement of single cell damage during photolysis under well defined physicochemical and photophysical conditions. Melanoma cells cultured in vitro were treated with the photosensitizer (PS), tin chlorin e6 (SnCe6) or immunoconjugate (SnCe6 conjugated to a anti-ICAM monoclonal antibody), and illuminated with a 10 mW He/Ne laser at a 630 nm wavelength. Cell membrane integrity was assessed using the vital dye calcein-AM. In experiments in which the laser power density and PS concentration were varied, it was determined that the time lag before cell rupture was inversely proportional to the estimated singlet oxygen flux to the cell surface. Microscopic examination of the lytic event indicated that photo-induced lysis was caused by a point rupture of the plasma membrane. The on-line nature of this microscopy system offers an opportunity to monitor the dynamics of the cell damage process and to gain insights into the mechanism governing photolytic cell injury processes.

Original languageEnglish (US)
Pages (from-to)2198-2206
Number of pages9
JournalBiophysical Journal
Volume68
Issue number5
DOIs
StatePublished - 1995

All Science Journal Classification (ASJC) codes

  • Biophysics

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