1 Introduction to the Detection of DNA Strand Breaks
7
Fig. 2. SSB detected by ISNT in human peripheral blood lymphocytes after UV irradiation
(Koji 1996). Human blood was drawn from
healthy male volunteers and separated in
Ficoll-Paque density-gradient centrifugation.
The lymphocyte fraction was collected and
washed with PBS. Finally, the PBL were suspended in 0.25 M sucrose. An aliquot of cell
suspension was irradiated with UV at a dose
of 10 J/m2, and then the cells were washed
with MEM. The final pellet was resuspended
in RPMI 1640 medium supplemented with
10% fetal bovine serum and allowed to repair
UV-induced DNA damage at 37° C under 5%
CO2 in air. At various time-points, an aliquot of
cell suspension was centrifuged onto gelatincoated glass slides, as detailed previously
(Koji 1996). The sites of SSB were detected
by ISNT as black spots. For more details, see
Chapter 2.
Oh
2h
24h
7
Fig. 2. SSB detected by ISNT in human peripheral blood lymphocytes after UV irradiation
(Koji 1996). Human blood was drawn from
healthy male volunteers and separated in
Ficoll-Paque density-gradient centrifugation.
The lymphocyte fraction was collected and
washed with PBS. Finally, the PBL were suspended in 0.25 M sucrose. An aliquot of cell
suspension was irradiated with UV at a dose
of 10 J/m2, and then the cells were washed
with MEM. The final pellet was resuspended
in RPMI 1640 medium supplemented with
10% fetal bovine serum and allowed to repair
UV-induced DNA damage at 37° C under 5%
CO2 in air. At various time-points, an aliquot of
cell suspension was centrifuged onto gelatincoated glass slides, as detailed previously
(Koji 1996). The sites of SSB were detected
by ISNT as black spots. For more details, see
Chapter 2.
Oh
2h
24h