FCS has never become established as a biophysical routine for particle sizing. This is due to the fact that sensitivity can, under certain conditions, indeed be disadvantageous, as it renders the technique error prone and overly susceptible to signal disturbances. Here, we discuss the systematic challenges, as well as the advances made over the past decades, to employing FCS in polydisperse samples. The problematic role of large particles, a common issue in DLS and FCS, and the effect of fluorescent labeling are discussed in detail, along with strategies for respective error mitigation in experiments and data analysis. We expect this overview to guide future users in successfully applying FCS to their particle sizing problems in the hope of fostering a more widespread and routine use of FCS-based technology.
FCS has never become established as a biophysical routine for particle sizing. This is due to the fact that sensitivity can, under certain conditions, indeed be disadvantageous, as it renders the technique error prone and overly susceptible to signal disturbances. Here, we discuss the systematic challenges, as well as the advances made over the past decades, to employing FCS in polydisperse samples. The problematic role of large particles, a common issue in DLS and FCS, and the effect of fluorescent labeling are discussed in detail, along with strategies for respective error mitigation in experiments and data analysis. We expect this overview to guide future users in successfully applying FCS to their particle sizing problems in the hope of fostering a more widespread and routine use of FCS-based technology.
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