Sample Preparation and Fluorescent Labeling

Fluorescent emission is required to visualize a macromolecule or structure by CSLM in fluorescent mode. In membrane apphcations, the fluorescent signal usually comes from the components of the filtrated solution and their fluorescent signal enables that they are visualized and located on the top or inside the membrane. 

The general requirements that a fluorescent probe has to fiilfill to be used in CSLM are the following  

The maximum absorption of the fluorescent probe has to be dose to the emission wavelength of the laser. 

The fluorescent emission must be intense (fluorescence intensity is directly proportional to the product of the molar extinction coefficient for absorption and the quantum yield for the fluorescence emission of the dye). 

Multilabeling experiments (in which more than one macromolecule or structure is stained) require fluorescent compounds whose excitation/emission wavelengths overlap minimally. 

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