The concepts of tail moment and tail inertia in the single cell gel electrophoresis assay.

Single cell gel electrophoresis under alkaline conditions is a technique used to detect primary DNA damage in individual mammalian cells. Cells embedded in agarose on microscope slides are subjected to lysis, unwinding of DNA and electrophoresis at high pH. After staining with a fluorescent dye, cells with DNA damage display increased migration of genetic material from the cell nucleus. The damage is quantified by measuring the displacement between the genetic material of the nucleus ('comet head') and the resulting 'tail'. The torsional moment of the tail ('tail moment') has been suggested to be an appropriate index of induced DNA damage in considering both the migration of the genetic material as well as the relative amount of DNA in the tail. In the present paper it will be shown that the moment of inertia ('tail inertia'), a not previously described tail parameter, provides a more precise description of the distribution of individual DNA fragments within the tails. The tail inertia was also found to be the most sensitive indicator of the DNA damage induced in peripheral lymphocytes from mice given a single intraperitoneal injection of cyclophosphamide (150 mg/kg b.w.). It is concluded that the tail inertia is an important complement to other tail parameters when looking for damage of DNA with the single cell gel electrophoresis assay.

[1]  R R Tice,et al.  Abundant alkali-sensitive sites in DNA of human and mouse sperm. , 1989, Experimental cell research.

[2]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[3]  R R Tice,et al.  DNA damage and repair with age in individual human lymphocytes. , 1990, Mutation research.

[4]  K J Johanson,et al.  Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. , 1984, Biochemical and biophysical research communications.

[5]  K. O’Neill,et al.  Application of confocal laser scanning microscopy to analysis of H2O2-induced DNA damage in human cells. , 1993, Scanning.

[6]  R E Durand,et al.  Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the "comet" assay. , 1990, Radiation research.

[7]  R. Tice,et al.  The single cell gel (SCG) assay: an electrophoretic technique for the detection of DNA damage in individual cells. , 1991, Advances in experimental medicine and biology.

[8]  M. Green,et al.  The single cell gel electrophoresis assay (comet assay): a European review. , 1993, Mutation research.

[9]  B. Hellman,et al.  Demonstration of chlorobenzene-induced DNA damage in mouse lymphocytes using the single cell gel electrophoresis assay. , 1995, Toxicology.

[10]  R. Jostes,et al.  Single-cell gel technique supports hit probability calculations. , 1993, Health physics.

[11]  R. Tice,et al.  High-dose combination alkylating agents with autologous bone-marrow support in patients with breast cancer: preliminary assessment of DNA damage in individual peripheral blood lymphocytes using the single cell gel electrophoresis assay. , 1992, Mutation research.