Abstract: A positioning and alignment instrument, and methods of use thereof, are provided for facilitating the alignment and insertion of a device, such as a guide wire, into bone. The instrument includes a handheld anchoring component and a rotatable guidance component. During use, the anchoring component is anchored into bone via anchoring protrusions, such that the position and orientation of the anchoring component is fixed relative to the bone. The guidance component, which is mechanically supported by the anchoring component, includes a device guide channel for receiving the device and guiding the device towards an insertion location adjacent to the distal end of the anchoring component. The guidance component is rotatable relative to the anchoring component about a rotation axis that is located adjacent to the distal end of the anchoring component, such that the insertion location remains adjacent to the distal end of the anchoring component under rotation.