Flow Cytometric Technique for Quantitating Cytotoxic Response to Photodynamic Therapy

Abstract— A simple flow cytometric technique for rapid measurement of multilog cytotoxic responses to photosensitization of cellular systems is described. This technique is particularly useful for cell lines with a low colony‐forming efficiency, for which a nonclonogenic assay is required. The assay separates cell‐sized objects from cellular debris by gating on forward scatter versus side scatter, identifies viable cells by positive calcein AM and negative ethidium homodimer‐1 staining and measures cell concentration relative to an internal standard of polystyrene beads. Large numbers of cells can be analyzed rapidly.

[1]  J A Steinkamp,et al.  Quantitation of cell concentration using the flow cytometer. , 2005, Cytometry.

[2]  R. Pieters,et al.  Mononuclear cells contaminating acute lymphoblastic leukaemic samples tested for cellular drug resistance using the methyl-thiazol-tetrazolium assay. , 1994, British Journal of Cancer.

[3]  J. Kennedy,et al.  Using δ-Aminolevulinic Acid in Cancer Therapy , 1994 .

[4]  S. Iinuma,et al.  A mechanistic study of cellular photodestruction with 5-aminolaevulinic acid-induced porphyrin. , 1994, British Journal of Cancer.

[5]  T. Pohl,et al.  Rapid quantification of lymphocyte subsets in heterogeneous cell populations by flow cytometry. , 1994, Cytometry.

[6]  H. Baker,et al.  Do glutathione and related enzymes play a role in drug resistance in small cell lung cancer cell lines? , 1993, British Journal of Cancer.

[7]  J. Kennedy,et al.  Endogenous protoporphyrin IX, a clinically useful photosensitizer for photodynamic therapy. , 1992, Journal of photochemistry and photobiology. B, Biology.

[8]  B. Wilson,et al.  RESISTANCE TO PHOTODYNAMIC THERAPY IN RADIATION INDUCED FIBROSARCOMA‐1 and CHINESE HAMSTER OVARY‐MULTI‐DRUG RESISTANT CELLS in vitro , 1991, Photochemistry and photobiology.

[9]  H. Pass,et al.  Sensitivity of different human lung cancer histologies to photodynamic therapy. , 1990, Cancer research.

[10]  L. Mchale,et al.  Use of a tetrazolium based colorimetric assay in assessing photoradiation therapy in vitro. , 1988, Cancer letters.

[11]  Richard P. Haugland,et al.  Handbook of fluorescent probes and research chemicals , 1996 .

[12]  I. Tannock,et al.  Flow cytometric analysis of doxorubicin accumulation in cells from human and rodent cell lines. , 1989, Journal of the National Cancer Institute.

[13]  S. Cole,et al.  Use of the MTT assay for rapid determination of chemosensitivity of human leukemic blast cells. , 1988, Leukemia research.

[14]  L. Weisenthal,et al.  Clonogenic and nonclonogenic in vitro chemosensitivity assays. , 1985, Cancer treatment reports.

[15]  L. Weisenthal,et al.  Comparison of dye exclusion assays with a clonogenic assay in the determination of drug-induced cytotoxicity. , 1983, Cancer research.