A robust method to measure viscosity of microquantities of biological samples, such as blood and mucus, could lead to a better understanding and diagnosis of diseases. Microsamples have presented persistent challenges to conventional rheology, which requires bulk quantities of a sample. Alternatively, fluid viscosity can be probed by monitoring microscale motion of particles. Here, we present a decorrelation-based method using M-mode phase-sensitive optical coherence tomography (OCT) to measure particle Brownian motion. This is similar to previous methods using laser speckle decorrelation but with sensitivity to nanometer-scale displacement. This allows for the measurement of decorrelation in less than 1 millisecond and significantly decreases sensitivity to bulk motion, thereby potentially enabling in vivo and in situ applications. From first principles, an analytical method is established using M-mode images obtained from a 47 kHz spectral-domain OCT system. A g(1) first-order autocorrelation is calculated from windows containing several pixels over a time frame of 200-1000 microseconds. Total imaging time is 500 milliseconds for averaging purposes. The autocorrelation coefficient over this short time frame decreases linearly and at a rate proportional to the diffusion constant of the particles, allowing viscosity to be calculated. In verification experiments using phantoms of microbeads in 200 µL glycerol-water mixtures, this method showed insensitivity to 2 mm/s lateral bulk motion and accurate viscosity measurements over a depth of 400 µm. In addition, the method measured a significant decrease of the apparent diffusion constant of soft tissue after formalin fixation, suggesting potential applications in mapping tissue stiffness.
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