Determination of vessel cross-sectional area by thresholding in Radon space.

The cross-sectional area of a blood vessel determines its resistance, and thus is a regulator of local blood flow. However, the cross-sections of penetrating vessels in the cortex can be non-circular, and dilation and constriction can change the shape of the vessels. We show that observed vessel shape changes can introduce large errors in flux calculations when using a single diameter measurement. Because of these shape changes, typical diameter measurement approaches, such as the full-width at half-maximum (FWHM) that depend on a single diameter axis will generate erroneous results, especially when calculating flux. Here, we present an automated method--thresholding in Radon space (TiRS)--for determining the cross-sectional area of a convex object, such as a penetrating vessel observed with two-photon laser scanning microscopy (2PLSM). The thresholded image is transformed back to image space and contiguous pixels are segmented. The TiRS method is analogous to taking the FWHM across multiple axes and is more robust to noise and shape changes than FWHM and thresholding methods. We demonstrate the superior precision of the TiRS method with in vivo 2PLSM measurements of vessel diameter.