In vivo visualization method by absolute blood flow velocity based on speckle and fringe pattern using two-beam multipoint laser Doppler velocimetry

Two-beam multipoint laser Doppler velocimetry (two-beam MLDV) is a non-invasive imaging technique able to provide an image of two-dimensional blood flow and has potential for observing cancer as previously demonstrated in a mouse model. In two-beam MLDV, the blood flow velocity can be estimated from red blood cells passing through a fringe pattern generated in the skin. The fringe pattern is created at the intersection of two beams in conventional LDV and two-beam MLDV. Being able to choose the depth position is an advantage of two-beam MLDV, and the position of a blood vessel can be identified in a three-dimensional space using this technique. Initially, we observed the fringe pattern in the skin, and the undeveloped or developed speckle pattern generated in a deeper position of the skin. The validity of the absolute velocity value detected by two-beam MLDV was verified while changing the number of layers of skin around a transparent flow channel. The absolute velocity value independent of direction was detected using the developed speckle pattern, which is created by the skin construct and two beams in the flow channel. Finally, we showed the relationship between the signal intensity and the fringe pattern, undeveloped speckle, or developed speckle pattern based on the skin depth. The Doppler signals were not detected at deeper positions in the skin, which qualitatively indicates the depth limit for two-beam MLDV.