Recognizing local artifacts in two-photon imaging of dendrites beneath blood vessels in vivo

Localized wavefront aberrations would introduce artifacts in biomedical imaging, which, however, are often neglected, as their compensations are at the cost of the field-of-view. Here, we show rarely reported local artifacts in two-photon imaging of dendrites beneath blood vessels in a mouse brain in vivo and interpret the phenomena via numerical simulations. The artifacts of divided parallel structures are found to be induced by coma and astigmatism, resulting from sample tilting and the cylinder shape of vasculatures, respectively. Different from that in single-photon microscopy, such artifacts in nonlinear microscopy show unique characteristics and should be recognized for proper interpretation of the images.