Monte Carlo simulation of fluorescence imaging of microvasculature



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Little numerical analysis has been done on fluorescence lifetime imaging \emph{in-vivo}. Here, a 3D fluorescence Monte Carlo model is used to evaluate a microvasculature geometry obtained via two-photon microscopy. I found that a bulk-vascularization assumption does not provide an accurate picture of penetration depth of the collected fluorescence signal. Instead the degree of absorption difference between extravascular and intravascular space, as well as the absorption difference between excitation and emission wavelengths must be taken into account to determine the depth distribution. Additionally, I found that using targeted illumination can provide for superior surface vessel sensitivity over wide-field illumination, with small area detection offering an even greater amount of sensitivity to surface vasculature. Depth sensitivity can be enhanced by either increasing the detector area or increasing the illumination area. Finally, it is shown that the excitation wavelength and vessel size can affect intra-vessel sampling distribution, as well as the amount of signal that originates from inside the vessel under targeted illumination conditions.