Multimodal vessel mapping for precise large area alignment of functional optical imaging data to neuroanatomical preparations in marmosets

Imaging technologies, such as intrinsic optical imaging (IOI), functional magnetic resonance imaging (fMRI) or multiphoton microscopy provide excellent opportunities to study the relationship between functional signals recorded from a cortical area and the underlying anatomical structure. This, in turn, requires accurate alignment of the recorded functional imaging data with histological datasets from the imaged tissue obtained after the functional experiment. This alignment is complicated by distortions of the tissue which naturally occur during histological treatment, and is particularly difficult to achieve over large cortical areas, such as primate visual areas. We present here a method that uses IOI vessel maps revealed in the time course of the intrinsic signal, in combination with vascular casts and vascular lumen labeling techniques together with a pseudo three dimensional (p3D) reconstruction of the tissue architecture in order to facilitate alignment of IOI data with posthoc histological datasets. We demonstrate that by such a multimodal vessel mapping approach, we are able to constitute a hook in anatomical-functional data alignment that enables the accurate assignment of functional signals over large cortical regions. As an example, we present precise alignments of IOI responses showing orientation selectivity of primate V1 with anatomical sections stained for cytochrome-oxidase-reactivity.