A spectral/Fourier domain optical coherence tomography (OCT) intravital microscope using a supercontinuum light source at 1.7 μm was developed to study subcortical structures noninvasively in the living mouse brain. The benefits of 1.7 μm for deep tissue brain imaging are demonstrated by quantitatively comparing OCT signal attenuation characteristics of cortical tissue across visible and near-infrared wavelengths. Imaging of hippocampal tissue architecture and white matter microvasculature are demonstrated in vivo through thinned-skull, glass coverslip-reinforced cranial windows in mice. Applications of this novel platform include monitoring disease progression and pathophysiology in rodent models of Alzheimer’s disease and subcortical dementias, including vascular dementia.

Quantification of chromophore concentrations in reflectance mode remains a major challenge for biomedical optics. Spectroscopic Optical Coherence Tomography (SOCT) provides depth-resolved spectroscopic information necessary for quantitative analysis of chromophores, like hemoglobin, but conventional SOCT analysis methods are applicable only to well-defined specular reflections, which may be absent in highly scattering biological tissue. Here, by fitting of the dynamic scattering signal spectrum in the OCT angiogram using a forward model of light propagation, we quantitatively determine hemoglobin concentrations directly. Importantly, this methodology enables mapping of both oxygen saturation and total hemoglobin concentration, or alternatively, oxyhemoglobin and deoxyhemoglobin concentration, simultaneously. Quantification was verified by ex vivo blood measurements at various pO₂ and hematocrit levels. Imaging results from the rodent brain and retina are presented. Confounds including noise and scattering, as well as potential clinical applications, are discussed.

The high acquisition speed of state-of-the-art optical coherence tomography (OCT) enables massive signal-to-noise ratio (SNR) improvements by signal averaging. Here, we investigate the performance of two commonly used approaches for OCT signal averaging. We present the theoretical SNR performance of (a) computing the average of OCT magnitude data and (b) averaging the complex phasors, and substantiate our findings with simulations and experimentally acquired OCT data. We show that the achieved SNR performance strongly depends on both the SNR of the input signals and the number of averaged signals when the signal bias caused by the noise floor is not accounted for. Therefore we also explore the SNR for the two averaging approaches after correcting for the noise bias and, provided that the phases of the phasors are accurately aligned prior to averaging, then find that complex phasor averaging always leads to higher SNR than magnitude averaging.

A method of measuring cortical oxygen metabolism in the mouse brain that uses independent quantitative measurements of three key parameters: cerebral blood flow (CBF), arteriovenous oxygen extraction (OE), and hemoglobin concentration ([HbT]) is presented. Measurements were performed using a single visible light spectral/Fourier domain OCT microscope, with Doppler and spectroscopic capabilities, through a thinned-skull cranial window in the mouse brain. Baseline metabolic measurements in mice are shown to be consistent with literature values. Oxygen consumption, as measured by this method, did not change substantially during minor changes either in the fraction of inspired oxygen (FiO₂) or in the fraction of inspired carbon dioxide (FiCO₂), in spite of larger variations in oxygen saturations. This set of experiments supports, but does not prove, the validity of the proposed method of measuring brain oxygen metabolism.

Optical Coherence Tomography Angiography (OCTA) refers to a powerful class of OCT scanning protocols and algorithms that selectively enhance the imaging of blood vessel lumens, based mainly on the motion and scattering of red blood cells (RBCs). Though OCTA is widely used in clinical and basic science applications for visualization of perfused blood vessels, OCTA is still primarily a qualitative tool. However, more quantitative hemodynamic information would better delineate disease mechanisms, and potentially improve the sensitivity for detecting early stages of disease. Here, we take a broader view of OCTA in the context of microvascular hemodynamics and light scattering. Paying particular attention to the unique challenges presented by capillaries versus larger supplying and draining vessels, we critically assess opportunities and challenges in making OCTA a quantitative tool.