I would like to examine the concentration of bio molecules (particularly glycoproteins) by using a fluorescent dye immobilizing these molecules on a membrane. I would like to mention that I have already performed this experiment in solution state and have measured the fluorescence intensity by using spectrofluorimeter quite easily. However, now I wish to do the same on a filter or nitrocellulose membrane.
We have not tried to measure fluorescence from a nitrocellulose membrane. To measure the fluorescence of samples run on gels, we scan the wet gel using a Typhoon scanner.
Reading fluorescence signal from membranes is usually done by fluorescence scanners (Typhoon Trio, Li-COR Odyssey etc) or you can also use high-end fluorescence readers (PheraSTAR, Synergy Neo) with scanning capability.
You can also try using front-face fluorescence if your spectrofluorimeter allows that. Some instruments - such as the ISS PC - allows one to utilize a variable angle front-face attachment. Otherwise it may be possible to jury-rig a piece of filter or nitrocellulose membrane in the sample compartment such that one can excite and observe the surface. This method is common for highly absorbing samples.
Dear All, I truly thank all of yours constructive suggestions. Yours scientific inputs have given me lot of confidence to go ahead in the direction to analyse my samples using fluorophores on the membrane. Best wishes
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