Autoradiography of Gels and Blots

There are two problems with autoradiography of gels (Laskey 1980). 

Isotopes such as or C emit low-energetic P-radiation. TTiis radiation is already absorbed in the gel and does not reach the film. 

Isotopes such as or 1 emit high-energetic P-radiation or y-radiation. This penetrates the film without blackening it. 

Fluorography (see Table 1.3) solves both problems. In case 1, the experimenter soaks the gel with a scintillator liquid (Enhance, Enlightning, Entensify). This converts the P-radiation into light. The light is not absorbed in the gel and thus reaches the film. However, the gels should not be stained with Coomassie Blue because the stain partially absorbs the light. Fluorography is also applicable to nitrocellulose blots. You spray the blot with Enhance Spray by NEN. 

For a H/ C fluorogram, the gel is fixed (without Coomassie ) and washed three times with 40% methanol and 10% acetic acid (15 minutes each), and then incubated for 15 to 30 minutes in a scintillator (ENTRANCE or ENLIGHTENING of NEN) and afterward dehydrated. ENLIGHTENING and EN HANCE are aggressive liquids. 

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