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Counting Samples from TLC Plates by LSC

In a typical TLC experiment, the radioactivity is detected at two points: after TLC it is analyzed by autoradiography, to locate radioactive spots. These spots are then scraped off of the plate and counted to provide quantitative information. Each of these steps can be enhanced using the following protocols.

Autoradiography and LSC with TLC Plates

  1. Autoradiography (Fluorography)


  2. After the plate has been developed, spray twice with National Diagnostics' Autofluor and allow the plate to dry. This impregnates the plate with phosphors, which will convert the b emissions to more readily detectable photons. Autofluor enhances the speed and sensitivity of detection when the plate is placed on film. Follow this link for details on autoradiography procedures.

  3. Liquid scintillation counting of scraped TLC silica


  4. Option 1: Suspend the silica powder in 10ml of a cocktail which has a gel phase with water such as National Diagnostics' Hydrofluor. Shake well, and add 3ml H2O. Shake until gel forms, and count.

    Option 2: Suspend the silica in 10ml of an organic based (non-emulsion) cocktail such as National Diagnostics' Ecoscint O. Add 0.5-1g finely divided silica thixotrophic agent to form a clear gel in the solution, keeping the TLC particles suspended. Count as usual.




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Products Related to this Discussion:
Autofluor
Water soluble scintillation phosphor for use as an autoradiographic image intensifier.
Hydrofluor
High efficiency, ready-to-use liquid scintillation solution designed for aqueous samples.
Ecoscint Ultra
Biodegradable scintillation cocktail with the best combination of sample hold and efficiency on the market. Ideal for low background counting of environmental samples.

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