Bertan Hallacoglu

Improvement of axial resolution and contrast in temporally focused widefield two-photon microscopy with structured light illumination

Although temporally focused wide-field two-photon microscopy (TFM) can perform depth resolved wide field imaging, it cannot avoid the image degradation due to scattering of excitation and emission photons when imaging in a turbid medium. Further, its axial resolution is inferior to standard point-scanning two-photon microscopy. We implemented a structured light illumination for TFM and have shown that it can effectively reject the out-of-focus scattered emission photons improving image contrast. Further, the depth resolution of the improved system is dictated by the spatial frequency of the structure light with the potential of attaining depth resolution better than point-scanning two-photon microscopy.

Absolute measurement of cerebral optical coefficients, hemoglobin concentration and oxygen saturation in old and young adults with near-infrared spectroscopy

We present near-infrared spectroscopy measurement of absolute cerebral hemoglobin concentration and saturation in a large sample of 36 healthy elderly (mean age, 85 ± 6 years) and 19 young adults (mean age, 28 ± 4 years). Non-invasive measurements were obtained on the forehead using a commercially available multi-distance frequency-domain system and analyzed using a diffusion theory model for a semi-infinite, homogeneous medium with semi-infinite boundary conditions. Our study included repeat measurements, taken five months apart, on 16 elderly volunteers that demonstrate intra-subject reproducibility of the absolute measurements with cross-correlation coefficients of 0.9 for absorption coefficient (μa), oxy-hemoglobin concentration ([HbO2]), and total hemoglobin concentration ([HbT]), 0.7 for deoxy-hemoglobin concentration ([Hb]), 0.8 for hemoglobin oxygen saturation (StO2), and 0.7 for reduced scattering coefficient (μs). We found significant differences between the two age groups. Compared to young subjects, elderly subjects had lower cerebral [HbO2], [Hb], [HbT], and StO2 by 10 ± 4 μM, 4 ± 3 μM, 14 ± 5 μM, and 6%±5%, respectively. Our results demonstrate the reliability and robustness of multi-distance near-infrared spectroscopy measurements based on a homogeneous model in the human forehead on a large sample of human subjects. Absolute, non-invasive optical measurements on the brain, such as those presented here, can significantly advance the development of NIRS technology as a tool for monitoring resting/basal cerebral perfusion, hemodynamics, oxygenation, and metabolism.

Practical steps for applying a new dynamic model to near-infrared spectroscopy measurements of hemodynamic oscillations and transient changes: implications for cerebrovascular and functional brain studies.

RATIONALE AND OBJECTIVES Perturbations in cerebral blood volume (CBV), blood flow (CBF), and metabolic rate of oxygen (CMRO2) lead to associated changes in tissue concentrations of oxy- and deoxy-hemoglobin (ΔO and ΔD), which can be measured by near-infrared spectroscopy (NIRS). A novel hemodynamic model has been introduced to relate physiological perturbations and measured quantities. We seek to use this model to determine functional traces of cbv(t) and cbf(t) - cmro2(t) from time-varying NIRS data, and cerebrovascular physiological parameters from oscillatory NIRS data (lowercase letters denote the relative changes in CBV, CBF, and CMRO2 with respect to baseline). Such a practical implementation of a quantitative hemodynamic model is an important step toward the clinical translation of NIRS. MATERIALS AND METHODS In the time domain, we have simulated O(t) and D(t) traces induced by cerebral activation. In the frequency domain, we have performed a new analysis of frequency-resolved measurements of cerebral hemodynamic oscillations during a paced breathing paradigm. RESULTS We have demonstrated that cbv(t) and cbf(t) - cmro2(t) can be reliably obtained from O(t) and D(t) using the model, and that the functional NIRS signals are delayed with respect to cbf(t) - cmro2(t) as a result of the blood transit time in the microvasculature. In the frequency domain, we have identified physiological parameters (e.g., blood transit time, cutoff frequency of autoregulation) that can be measured by frequency-resolved measurements of hemodynamic oscillations. CONCLUSIONS The ability to perform noninvasive measurements of cerebrovascular parameters has far-reaching clinical implications. Functional brain studies rely on measurements of CBV, CBF, and CMRO2, whereas the diagnosis and assessment of neurovascular disorders, traumatic brain injury, and stroke would benefit from measurements of local cerebral hemodynamics and autoregulation.

Validation of a novel hemodynamic model for coherent hemodynamics spectroscopy (CHS) and functional brain studies with fNIRS and fMRI

We report an experimental validation and applications of the new hemodynamic model presented in the companion article (Fantini, 2014-this issue) both in the frequency domain and in the time domain. In the frequency domain, we have performed diffuse optical measurements for coherent hemodynamics spectroscopy (CHS) on the brain and calf muscle of human subjects, showing that the hemodynamic model predictions (both in terms of spectral shapes and absolute spectral values) are confirmed experimentally. We show how the quantitative analysis based on the new model allows for autoregulation measurements from brain data, and provides an analytical description of near-infrared spiroximetry from muscle data. In the time domain, we have used data from the literature to perform a comparison between brain activation signals measured with functional near-infrared spectroscopy (fNIRS) or with blood oxygenation level dependent (BOLD) fMRI, and the corresponding signals predicted by the new model. This comparison shows an excellent agreement between the model predictions and the reported fNIRS and BOLD fMRI signals. This new hemodynamic model provides a valuable tool for brain studies with hemodynamic-based techniques.

Reduced speed of microvascular blood flow in hemodialysis patients versus healthy controls: a coherent hemodynamics spectroscopy study

Abstract. We present a pilot clinical application of coherent hemodynamics spectroscopy (CHS), a technique to investigate cerebral hemodynamics at the microcirculatory level. CHS relies on frequency-resolved measurements of induced cerebral hemodynamic oscillations that are measured with near-infrared spectroscopy (NIRS) and analyzed with a hemodynamic model. We have used cyclic inflation (200 mmHg) and deflation of a pneumatic cuff placed around the subject’s thigh at seven frequencies in the range of 0.03 to 0.17 Hz to generate CHS spectra and to obtain a set of physiological parameters that include the blood transit times in the cerebral microcirculation, the cutoff frequency for cerebral autoregulation, and blood volume ratios across the three different compartments. We have investigated five hemodialysis patients, during the hemodialysis procedure, and six healthy subjects. We have found that the blood transit time in the cerebral microcirculation is significantly longer in hemodialysis patients with respect to healthy subjects. No significant differences were observed between the two groups in terms of autoregulation efficiency and blood volume ratios. The demonstration of the applicability of CHS in a clinical setting and its sensitivity to the highly important cerebral microcirculation may open up new opportunities for NIRS applications in research and in medical diagnostics and monitoring.

Cerebral perfusion and oxygenation are impaired by folate deficiency in rat: absolute measurements with noninvasive near-infrared spectroscopy

Brain microvascular pathology is a common finding in Alzheimers disease and other dementias. However, the extent to which microvascular abnormalities cause or contribute to cognitive impairment is unclear. Noninvasive near-infrared spectroscopy (NIRS) can address this question, but its use for clarifying the role of microvascular dysfunction in dementia has been limited due to theoretical and practical considerations. We developed a new noninvasive NIRS method to obtain quantitative, dynamic measurements of absolute brain hemoglobin concentration and oxygen saturation and used it to show significant cerebrovascular impairments in a rat model of diet-induced vascular cognitive impairment. We fed young rats folate-deficient (FD) and control diets and measured absolute brain hemoglobin and hemodynamic parameters at rest and during transient mild hypoxia and hypercapnia. With respect to control animals, FD rats featured significantly lower brain hemoglobin concentration (72±4 μmol/L versus 95±6 μmol/L) and oxygen saturation (54%±3% versus 65%±2%). By contrast, resting arterial oxygen saturation was the same for both groups (96%±4%), indicating that decrements in brain hemoglobin oxygenation were independent of blood oxygen carrying capacity. Vasomotor reactivity in response to hypercapnia was also impaired in FD rats. Our results implicate microvascular abnormality and diminished oxygen delivery as a mechanism of cognitive impairment.

Noninvasive assessment of testicular torsion in rabbits using frequency-domain near-infrared spectroscopy: prospects for pediatric urology

We present a quantitative near-IR spectroscopy study of the absolute values of oxygen saturation of hemoglobin before and after surgically induced testicular torsion in adult rabbits. Unilateral testicular torsions (0, 540, or 720 deg) on experimental testes and contralateral sham surgery on control testes are performed in four adult rabbits. A specially designed optical probe for measurements at multiple source-detector distances and a commercial frequency-domain tissue spectrometer are used to measure absolute values of testicular hemoglobin saturation. Our results show: (1) a consistent baseline absolute tissue hemoglobin saturation value of 78+/-5%, (2) a comparable tissue hemoglobin saturation of 77+/-6% after sham surgery, and (3) a significantly lower tissue hemoglobin saturation of 36+/-2% after 540- and 720-deg testicular torsion surgery. Our findings demonstrate the feasibility of performing frequency-domain, multidistance near-IR spectroscopy for absolute testicular oximetry in the assessment of testicular torsion. We conclude that near-IR spectroscopy has potential to serve as a clinical diagnostic and monitoring tool for the assessment of absolute testicular hemoglobin desaturation caused by torsion, with the possibility of serving as a complement to conventional color and spectral Doppler ultrasonography.

Cerebral Blood Volume and Vasodilation are Independently Diminished by Aging and Hypertension: A Near Infrared Spectroscopy Study

BACKGROUND Senescent changes in brain microvascular circulation may cause or contribute to age-related cognitive decline. Such changes are promoted partly by aging, but also by chronic hypertension, a leading treatable cause of cognitive decline. OBJECTIVES We aimed to non-invasively detect in vivo the senescent changes in brain microvascular circulation associated with age and hypertension, and inquired whether decrements driven by aging would be exacerbated by chronic hypertension. METHODS In this longitudinal study, absolute near infrared spectroscopy (NIRS) was used to quantify in vivo cerebral blood volume (CBV) and assess the hemodynamic response to a hypercapnic respiratory challenge in normotensive Wistar-Kyoto (WKY) and spontaneous-hypertensive (SHR) rats. The impact of age and hypertension were evaluated by repeating these measurements on the same animals at 4- and 16-months of age. RESULTS CBV decreased markedly with age in both strains, from 4.5 ± 0.2 to 2.6 ± 0.1 ml/100g tissue, on average. Chronic hypertension, however, did not significantly exacerbate this age-related decrease in CBV (-48.1 ± 3.7% in WKYs versus -53.3 ± 5.4% in SHRs). In contrast, vasoreactivity was already impaired in the young hypertensive rats (ΔVMR 0.017 ± 0.014 in young SHRs versus 0.042 ± 0.005 in young WKYs) and further worsened by middle-age (ΔVMR 0.011 ± 0.017 middle-aged SHRs). CONCLUSION Whereas a decrease in brain blood volume correlated with age but not hypertension, vasodilatory capacity was diminished due to hypertension but did not appear affected by age alone. The ability of absolute NIRS to distinguish between such senescent changes in brain (micro)vascular circulation in life may allow early detection and intervention to preserve cerebrovascular health with age.

COHERENT HEMODYNAMICS SPECTROSCOPY BASED ON A PACED BREATHING PARADIGM — REVISITED

A novel hemodynamic model has been recently introduced, which provides analytical relationships between the changes in cerebral blood volume (CBV), cerebral blood flow (CBF), and cerebral metabolic rate of oxygen (CMRO2), and associated changes in the tissue concentrations of oxy- and deoxy-hemoglobin (ΔO and ΔD) measured with near-infrared spectroscopy (NIRS) [S. Fantini, Neuroimage85, 202–221 (2014)]. This novel model can be applied to measurements of the amplitude and phase of induced hemodynamic oscillations as a function of the frequency of oscillation, realizing the novel technique of coherent hemodynamics spectroscopy (CHS) [S. Fantini, Neuroimage85, 202–221 (2014); M. L. Pierro et al., Neuroimage85, 222–233 (2014)]. In a previous work, we have demonstrated an in vivo application of CHS on human subjects during paced breathing [M. L. Pierro et al., Neuroimage85, 222–233 (2014)]. In this work, we present a new analysis of the collected data during paced breathing based on a slightly revised formulation of the hemodynamic model and an efficient fitting procedure. While we have initially treated all 12 model parameters as independent, we have found that, in this new implementation of CHS, the number of independent parameters is eight. In this article, we identify the eight independent model parameters and we show that our previous results are consistent with the new formulation, once the individual parameters of the earlier analysis are combined into the new set of independent parameters.

Applications of a novel hemodynamic model to functional brain studies with fNIRS and fMRI

We report time-domain applications of a new hemodynamic model by Fantini [1] that yields analytic expressions for signals that are measurable with hemodynamic-based neuroimaging techniques such as functional near-infrared spectroscopy (fNIRS) and functional magnetic resonance imaging (fMRI). We show how the model can be used to predict the perturbations in cerebral blood volume (CBV), blood flow (CBF), and metabolic rate of oxygen (CMRO2) that account for the initial dip and post-stimulus undershoot that have been reported in the fMRI and fNIRS literature. Furthermore, we have used data from the literature to perform a comparison between measured fNIRS and fMRI signals and the corresponding signals predicted by the new hemodynamic model. Results showed an excellent agreement between the model predictions and the reported measured data.