1. Radiation Cell Survival Curves

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