Non-invasive fluorescence retinal imaging in small animals is an important requirement for an array of translational vision applications. The in vivo two-photon imaging of the mouse retina may enable the long-term investigation of the structure and function of healthy and diseased retinal tissue. However, to date, this has only been possible using relatively complex adaptive-optics systems. Here, the optical modeling of the murine eye and of the imaging system is used to achieve correction-free two-photon microscopy through the pupil of a mouse eye to yield high-quality, optically sectioned fundus images. By remotely scanning the focus using an electronically tunable lens, high-resolution three-dimensional fluorescein angiograms and cellular-scale images are acquired, thus introducing a correction-free baseline performance level for two-photon in vivo retinal imaging. Moreover, the system enables functional calcium imaging of repeated retinal responses to light stimulation using the genetically encoded indicator, GCaMP6s. These results and the simplicity of the new add-on optics are an important step toward several structural, functional, and multimodal imaging applications that will benefit from the tight optical sectioning and the use of near-infrared light.

Currently there is no accurate objective measure for monitoring pain during the state of drug-induced unconsciousness (such as during surgical anesthesia). Moreover, the absence of an objective measure for detecting pain hampers the physician's ability to provide an optimal dose of analgesics. We have developed a novel method for detecting pain by quantifying skin blood flow dynamics using a miniaturized dynamic light scattering (mDLS) sensor placed on the skin. Healthy awake volunteers were studied with mDLS sensors placed on both index fingers while being subjected to a series of cutaneous painful stimuli (electric shock and heat), randomly applied in a range between the subjects' pain threshold and tolerance. Power spectrum analysis of the recorded signal was performed with a focus on two frequency bands, representing relative blood flow of non-pulsatile vessels and larger pulsatile arterioles. Relative blood flow of pulsatile vessels decreased while flow of non-pulsatile vessels increased in response to painful stimulation, with a high correlation between the responses obtained on the right and left index fingers. The changes in hemodynamics that occur during painful stimulation suggest a redistribution of blood flow between pulsatile and non-pulsatile vessels, probably related to central activation of the sympathetic system combined with local dynamic autoregulatory responses. Thus, optical parameters of skin blood flow can detect nociceptive stimuli and consequently can serve as objective biomarkers of pain.

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