Standard Protocols Autoradiography (35S)

 

• Remove Kodak NTB2 nuclear emulsion from fridge and place at 42^o C for around 30-60 mins (until melted).

• Make up the developer and the fixer and place in the darkroom ready for later (only if developing the same day ). The developer should be a 1:1 dilution in water. The fixer is 30% (w/v) of sodium thisulphate.

• Place slides in a dark box and carry to dark room. Also take a slide rack, tweezers and the emulsion.

• Switch on the red light and switch off the main light.

• Working with the red light, unwrap the foil from the emulsion. Place the emulsion in the slide rack.

• As quick as possible dip a slide into the emulsion with the tweezers. Place the slide in the slide rack to dry. The quicker the slide is dipped, the thinner the coating of emulsion and the better the visualisation later.

• Switch off the red light.

• After around 30-60 minutes the slides will be dry. If you don’t want to wait in the dark for half an hour make sure to place a note on the darkroom so people know not to enter.

• After the slides are dry, re-enter the room but don’t turn on the light . Working in the dark, place the slides into the dark box and wrap in foil. Also wrap up the emulsion.

• Store slides in a fridge or cold room

 

Developing the slides:

 

• Remove slides from fridge and allow to warm to room temperature for around an hour

• Working in the dark, place the slides in a slide holder and immerse the slide holder in D19 developer from 3 minutes. Time with a countdown timer as you won’t be able to tell the time.

• Immediately place the slide holder in the fixer. Leave for 3 minutes again.

• Turn on the main light.

• Wash the slides with running tap water for around 5 minutes.