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Measuring key physiological parameters of small blood samples extracted from patients could be useful for real-time clinical diagnosis at the point of care. An important parameter required from all blood tests is the blood hematocrit, a measure of the fractional volume occupied by the red cells within the blood. In this work, we present a method for in vitro evaluation of hematocrit based on the data acquired using spectrally encoded flow cytometry. Analysis of the reflectance confocal images of blood within a flow chamber resulted in an error as low as 1.7% in the measured hematocrit. The technique could be used as part of an in vitro diagnostic system that measures important blood parameters at the point of care.

Spectrally encoded flow cytometry (SEFC) is a promising technique for noninvasive in vivo microscopy of blood cells. Here, we introduce a novel SEFC system for label-free confocal imaging of blood cells flowing at velocities of up to 10  mm/s within 65 μm-diameter vessels. The new system employs interferometric Fourier-domain detection and a high-speed wavelength-swept source, allowing 100 kHz line rate, sufficient for sampling the rapidly flowing cells 80 μm below the tissue surface. The large data sets obtained by this technique would improve diagnosis accuracy, reduce imaging time, and open new possibilities for noninvasive monitoring of blood in patients.

Optical microscopy of blood cells in vivo provides a unique opportunity for clinicians and researchers to visualize the morphology and dynamics of circulating cells, but is usually limited by the imaging speed and by the need for exogenous labeling of the cells. Here we present a label-free approach for in vivo flow cytometry of blood using a compact imaging probe that could be adapted for bedside real-time imaging of patients in clinical settings, and demonstrate subcellular resolution imaging of red and white blood cells flowing in the oral mucosa of a human volunteer. By analyzing the large data sets obtained by the system, valuable blood parameters could be extracted and used for direct, reliable assessment of patient physiology.

Holographic speckle is a major impediment for the emerging applications of multiphoton holographic projection in biomedical imaging, photo-stimulation and micromachining. Time averaging of multiple shifted versions of a single hologram ("shift-averaging") is a computationally-efficient method that was recently shown to deterministically eliminate holographic speckle in single-photon applications. Here, we extend these results and show, computationally and experimentally, that in two-photon holographic excitation shift-averaging also reduces holographic speckle better than "random" averaging of multiple calculated holograms.

Flow cytometry techniques often rely on detecting fluorescence from single cells flowing through the cross section of a laser beam, providing invaluable information on vast numbers of cells. Such techniques, however, are often limited in their ability to resolve clusters of cells or parallel cell flow through large vessels. We present a confocal imaging technique that images unstained cells flowing in parallel through a wide channel, using spectrally encoded reflectance confocal microscopy that does not require mechanical scanning. Images of red blood cells from our system are compared to conventional transmission microscopy, and imaging of flowing red blood cells in vitro is experimentally demonstrated.

Speckle is a major source of noise in holographic projection. Time averaging of multiple holograms may be used to reduce speckle contrast, but multiple holograms must be calculated per each frame, costing in computational power. We show that a single hologram may be used to generate a fully speckle-free reconstruction, by cyclic shifting and time averaging. We demonstrate the concept experimentally, and discuss its application for high-rate holographic projection systems.