Place the culture bottles upside down under an X-ray machine or 60Co source and shade the desired colonies with a lead block.

Operation method

Scheme 14.7 Separation of cell colonies by radiation method

Principle

Place the culture bottles upside down under an X-ray machine or 60Co source and shade the desired colonies with a lead block.

Materials and Instruments

D-PBS 0.25% trypsin
Growth medium X-ray or cobalt source Lead block

Move

1. Select the desired drop and mark it with a felt-tip pen or Nikon marker.

2. Select an appropriately sized piece of lead.

3. Hold the culture bottle to the radioactive source.

4. Place the culture bottle upside down under the source.

5. Shield the desired colony with a 2 mm thick lead block (a block of about 2 to 5 mm in diameter cut from a 2 mm thick strip of lead).

6. Irradiate the culture flask with 30 Gy of radiation.

7. Return the culture flasks to a sterile environment, remove the medium, trypsinize and re-grow the cells in the original culture flasks. Cells receiving radiation can be used as feeder layer cells.

Caveat

X-ray machine or60 Co source must be used under strict supervision.Co source should always be used under strict supervision and properly inspected to ensure the safety of exposed areas for yourself and others. Contact your local radiological protection officer before performing such tests.

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