A method to obtain reference images for evaluation of ultrasonic tissue characterization techniques

A general problem when evaluating ultrasonic methods for tissue characterization is that "a golden standard" is seldom known. This paper describes a manual method to obtain a reference image, with the same geometry as the ultrasound image, indicating spatial location of the different tissue types present in the biological tissue scanned in vitro. A 30 x 10 x 2 mm3 piece of formalin fixed porcine tissue was molded into an agar block, which on the top surface, contained a set of fiducial markers, spaced 2.5 mm. The block was submerged into 20 degrees C water and a set of parallel 7.5 MHz spatial compound ultrasound images of tissue and fiducial markers were recorded each 0.5 mm. Guided by the fiducial markers, the agar block was subsequently cut into slices 2.5 mm thick, photographed and finally analyzed histologically identifying these tissues: collagen rich, collagen poor, micro vessels and muscle fibres. Due to: (1) the cutting procedure, (2) the finite size of the ultrasound beam and (3) the spatial variation in propagation velocity, the macroscopic photographs did not align completely with the ultrasound images. Likewise, the histological image is a geometrically distorted version of the macroscopic photograph, due to the histological preparation process. The histological information was "mapped back" into the format of the ultrasound images the following way: On the macroscopic images, outlines were drawn manually which defined the border of the tissue. These outlines were superimposed on the corresponding ultrasound images (identified via the fiducial markers) and modified to encompass what appeared to be tissue regions on the ultrasound images and subsequently re-applied to the macroscopic image. This modified macroscopic outline was used as guideline when drawing outlines identifying regions of the various tissue types. Specifically, the macroscopic image revealed the borders between the different tissues, while the histological image identified the four tissue types. A set of 12 reference images based on modified macroscopic outlines was created. The overlap between the ultrasound images and the macroscopic images--which are the geometrical basis for the final reference images--was between 77% and 93%. A set of 12 reference images spaced 2.5 mm, identifying spatial location of four different tissue types in porcine muscle has been created. With the reference images, it is possible to quantitatively compare different ultrasound based tissue classification techniques.