Uma Maheswari Rajagopalan

Functional optical coherence tomography reveals localized layer-specific activations in cat primary visual cortex in vivo

Surface neural activity has been widely visualized using optical intrinsic signal imaging (OISI) from various cortical sensory areas. OISI of the cortical surface with a CCD camera gives integrated information across a depth of a few hundred micrometers. We visualize depth-resolved activation patterns of cat primary visual cortex by functional optical coherence tomography (fOCT). A comparison of the depth-integrated results of fOCT maps with the optical intrinsic signal profiles shows fairly good agreement. Our results reveal layer-specific activation patterns and indicate that the activation was not homogeneous.

In vivo layer visualization of rat olfactory bulb by a swept source optical coherence tomography and its confirmation through electrocoagulation and anatomy

Here, we report in vivo 3-D visualization of the layered organization of a rat olfactory bulb (OB) by a swept source optical coherence tomography (SS-OCT). The SS-OCT operates at a wavelength of 1334 nm with respective theoretical depth and lateral resolutions of 6.7 μm and 15.4 μm in air and hence it is possible to get a 3D structural map of OB in vivo at the micron level resolution with millimeter-scale imaging depth. Up until now, with methods such as MRI, confocal microscopy, OB depth structure in vivo had not been clearly visualized as these do not satisfy the criterion of simultaneously providing micron-scale spatial resolution and imaging up to a few millimeter in depth. In order to confirm the OB’s layered organization revealed by SS-OCT, we introduced the technique of electrocoagulation to make landmarks across the layered structure. To our knowledge this is such a first study that combines electrocoagulation and OCT in vivo of rat OB. Our results confirmed the layered organization of OB, and moreover the layers were clearly identified by electrocoagulation landmarks both in the OCT structural and anatomical slice images. We expect such a combined study is beneficial for both OCT and neuroscience fields.

Swept source optical coherence tomography as a tool for real time visualization and localization of electrodes used in electrophysiological studies of brain in vivo.

In studies of in vivo extracellular recording, we usually penetrate electrodes almost blindly into the neural tissue, in order to detect the neural activity from an expected target location at a certain depth. After the recording, it is necessary for us to determine the position of the electrodes precisely. Generally, to identify the position of the electrode, one method is to examine the postmortem tissue sample at micron resolution. The other method is using MRI and it does not have enough resolution to resolve the neural structures. To solve such problems, we propose swept source optical coherence tomography (SS-OCT) as a tool to visualize the cross-sectional image of the neural target structure along with the penetrating electrode. We focused on a rodent olfactory bulb (OB) as the target. We succeeded in imaging both the OB layer structure and the penetrating electrode, simultaneously. The method has the advantage of detecting the electrode shape and the position in real time, in vivo. These results indicate the possibility of using SS-OCT as a powerful tool for guiding the electrode into the target tissue precisely in real time and localizing the electrode tip during electrophysiological recordings.

Functional optical coherence tomography of rat olfactory bulb with periodic odor stimulation

In rodent olfactory bulb (OB), optical intrinsic signal imaging (OISI) is commonly used to investigate functional maps to odorant stimulations. However, in such studies, the spatial resolution in depth direction (z-axis) is lost because of the integration of light from different depths. To solve this problem, we propose functional optical coherence tomography (fOCT) with periodic stimulation and continuous recording. In fOCT experiments of in vivo rat OB, propionic acid and m-cresol were used as odor stimulus presentations. Such a periodic stimulation enabled us to detect the specific odor-responses from highly scattering brain tissue. Swept source OCT operating at a wavelength of 1334 nm and a frequency of 20 kHz, was employed with theoretical depth and lateral resolutions of 6.7 μm and 15.4 μm, respectively. We succeeded in visualizing 2D cross sectional fOCT map across the neural layer structure of OCT in vivo. The detected fOCT signals corresponded to a few glomeruli of the medial and lateral parts of dorsal OB. We also obtained 3D fOCT maps, which upon integration across z-axis agreed well with OISI results. We expect such an approach to open a window for investigating and possibly addressing toward inter/intra-layer connections at high resolutions in the future.

Using the Light Scattering Component of Optical Intrinsic Signals to Visualize In Vivo Functional Structures of Neural Tissues

Visualization of changes in reflected light from in vivo brain tissues reveals spatial patterns of neural activity. An important factor which influences the degree of light reflected includes the change in light scattering elicited by neural activation. Microstructures of neural tissues generally cause light scattering, and neural activities are associated with some changes in the microstructures. Here, we show that the optical properties unique to light scattering enable us to visualize spatial patterns of retinal activity non-invasively (FRG: functional retinography), and resolve functional structures in depth (fOCT: functional optical coherence tomography).

3D topology of orientation columns in visual cortex revealed by functional optical coherence tomography

Orientation tuning is a canonical neuronal response property of six-layer visual cortex that is encoded in pinwheel structures with center orientation singularities. Optical imaging of intrinsic signals enables us to map these surface two-dimensional (2D) structures, whereas lack of appropriate techniques has not allowed us to visualize depth structures of orientation coding. In the present study, we performed functional optical coherence tomography (fOCT), a technique capable of acquiring a 3D map of the intrinsic signals, to study the topology of orientation coding inside the cat visual cortex. With this technique, for the first time, we visualized columnar assemblies in orientation coding that had been predicted from electrophysiological recordings. In addition, we found that the columnar structures were largely distorted around pinwheel centers: center singularities were not rigid straight lines running perpendicularly to the cortical surface but formed twisted string-like structures inside the cortex that turned and extended horizontally through the cortex. Looping singularities were observed with their respective termini accessing the same cortical surface via clockwise and counterclockwise orientation pinwheels. These results suggest that a 3D topology of orientation coding cannot be fully anticipated from 2D surface measurements. Moreover, the findings demonstrate the utility of fOCT as an in vivo mesoscale imaging method for mapping functional response properties of cortex in the depth axis. NEW & NOTEWORTHY We used functional optical coherence tomography (fOCT) to visualize three-dimensional structure of the orientation columns with millimeter range and micrometer spatial resolution. We validated vertically elongated columnar structure in iso-orientation domains. The columnar structure was distorted around pinwheel centers. An orientation singularity formed a string with tortuous trajectories inside the cortex and connected clockwise and counterclockwise pinwheel centers in the surface orientation map. The results were confirmed by comparisons with conventional optical imaging and electrophysiological recordings.

Highly sensitive optical interferometric technique reveals stress-dependent instantaneous nanometric growth fluctuations of Chinese chive leaf under heavy metal stress

Plant growth apart from being a complex and highly dynamic is dependent on its immediate environment. Leaf expansion measurements using Statistical Interferometry Technique, a sensitive interferometric technique at nanometric accuracy and at sub-second levels revealed the presence of characteristic nanometric intrinsic fluctuations [Plant Biotechnology 31, 195 (2014)]. In this paper, we demonstrate that the nanometric intrinsic fluctuations are sensitive enough that they change under exposure of heavy metals, essential micronutrient zinc and non-essential element cadmium, at relatively low concentrations in the leaves of Chinese chive (Allium tuberosum). The nanometric intrinsic fluctuations of leaves were observed for 4h under three cadmium concentrations or two zinc concentrations. Results showed significant reduction of nanometric intrinsic fluctuations for all cadmium concentrations, and in contrast significant increase of nanometric intrinsic fluctuations for all zinc concentrations. There was significant reduction of nanometric intrinsic fluctuations for cadmium exposure of concentrations of 0.001mM for even an hour, and significant increment of nanometric intrinsic fluctuations under 0.75mM zinc from 1hr exposure. For comparison, antioxidative enzymes and metal uptake were also measured under 4hr exposure of cadmium or zinc. However, no significant changes could be seen in antioxidative enzymes within 4h under the smaller concentration of 0.001mM cadmium as seen for nanometric intrinsic fluctuations. The results imply that nanometric intrinsic fluctuations can be not only used as a measure for heavy metal stress but also it can be more sensitive to detect the toxic as well as positive effects of smaller amounts of heavy metal on plants at an early stage.

Monitoring of growth dynamics of plants under the influence of cadmium using a highly sensitive interferometry technique

Abstract. Using statistical interferometry technique (SIT), a highly sensitive interferometry technique developed in our laboratory, we reported about the existence of nanometric intrinsic fluctuations (NIF) in a variety of plants. SIT permits noncontact, noninvasive, and fast detection of plant growth fluctuations in subnanometer scale. We propose the application of NIF to investigate the effect of heavy metal, cadmium, on growth dynamics of Chinese chive (Allium tuberosum). NIFs of leaves were observed for 3 days under four different concentrations of CdCl2: 0, 0.001, 0.01, and 0.1 mM. Results showed significant reduction of NIFs within 4 h for all Cd concentrations, and there was a further decrease with the exposure time of Cd under 0.1 and 0.01 mM. In addition, under 0.001 mM, a significant recovery could be observed after a rapid reduction in the first 4 h. As a comparison, measured antioxidative enzymes increased with increasing Cd concentration. However, no significant increase could be seen within the initial 4 h under a smaller concentration of 0.001 mM as seen for NIFs. The results imply that NIF can be used as an indicator for heavy metal stress on plants as well as it can be more sensitive to detect the influence of smaller Cd amounts on plants at an early stage.

Functional imaging of cat primary visual cortex with optical coherence tomography

We report the application of Optical coherence tomography (OCT) for visualizing a one dimensional depth resolved functional structure of cat brain in vivo. The OCT system is based on the known fact that neural activation induces structural changes such as capillary dilation and cellular swelling. Detecting these changes as an amplitude change of the scattered light, an OCT signal reflecting neural activity i.e., fOCT (functional OCT) could be obtained. Experiments have been done to obtain a depth resolved stimulus-specific profile of activation in cat visual cortex. Our results in one dimension indicate that indeed an orientation dependent functional signal could be obtained. Further, we show that this depth resolved fOCT signal is well correlated with the stimulus dependent column determined by OISI. Based on the results, the smallest functional unit in depth, resolved by the proposed system is around 40 micrometers . We are extending our system to perform two dimensional functional imaging.