Embedded Files
Post date: Jul 9, 2012 4:33:58 AM
1.1 Aims
· Measure survival of cells after exposure to various x-ray doses.
· Determine the quantity, SF2 – Surviving Fraction after 2 Gy
1.2 Tools
· Cells in media
· 6-well plate or T25 flask
· X-ray source
· Cell Counter (Hemacytometer)
1.3 Methods
1.3.1 Cell Culture
· Seed at the following approximate concentrations of cells for each radiation dose:
o 300 cells for 0 Gy
o 600 cells for 2 Gy
o 1200 cells for 4 Gy
o 2400 cells for 6 Gy
o 4800 cells for 8 Gy
o 9600 cells for 10 Gy
· Seed cells either in T25ml flasks OR 6 well plates
· After seeding cover cell culture with media (5ml for T25 flask)
1.3.2 Irradiation
· Faxtron
· Linac
· Wait 8-10 days post irradiation for formation of colonies
1.3.3 Analysis
· Remove media
· Add 5 mL of 4% formalin (for T25 flasks)
· Leave for 2 hours, then remove the formalin
· Stain with 0.5% methylene blue (0.5g/100ml) by adding an amount that is sufficient to cover the plate
o Leave for 12-24 hours
· Remove methylene blue and count number of colonies
Report abuse