-
[show abstract]
[hide abstract]
ABSTRACT: We report the development of an all-fiber-optic scanning endomicroscope capable of high-resolution second harmonic generation (SHG) imaging of biological tissues and demonstrate its utility for monitoring the remodeling of cervical collagen during gestation in mice. The endomicroscope has an overall 2.0 mm diameter and consists of a single customized double-clad fiber, a compact rapid two-dimensional beam scanner, and a miniature compound objective lens for excitation beam delivery, scanning, focusing, and efficient SHG signal collection. Endomicroscopic SHG images of murine cervical tissue sections at different stages of normal pregnancy reveal progressive, quantifiable changes in cervical collagen morphology with resolution similar to that of bench-top SHG microscopy. SHG endomicroscopic imaging of ex vivo murine and human cervical tissues through intact epithelium has also been performed. Our findings demonstrate the feasibility of SHG endomicroscopy technology for staging normal pregnancy, and suggest its potential application as a minimally invasive tool for clinical assessment of abnormal cervical remodeling associated with preterm birth.
Proceedings of the National Academy of Sciences 07/2012; 109(32):12878-83. · 9.68 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report recent advances in ultrathin fiber-optic endomicroscopy technology with significant improvements on resolution and signal collection. Ex vivo and in vivo two-photon imaging of unstained biological tissues and potential clinical applications will be presented.
05/2012;
-
[show abstract]
[hide abstract]
ABSTRACT: We compare the illumination uniformity and the associated effects of the spiral and Lissajous scanning patterns that are commonly used in an endomicroscope. Theoretical analyses and numerical simulations were first performed to quantitatively investigate the area illumination density in the spiral scanning pattern. The results revealed the potential problem of manifest photodamage due to the very high illumination density in the center of the spiral scan. Similar analyses of the Lissajous scanning pattern, which can be conveniently implemented on the same endomicroscope with no hardware modifications, showed a more uniform illumination density with about an 80-fold reduction in the peak illumination density. To underscore the benefit offered by the improved illumination uniformity, we conducted in vitro two-photon fluorescence imaging of cultured cells stained with a LIVE/DEAD viability assay using our home-built, fiber-optic, two-channel endomicroscopy system. Both the spiral and the Lissajous scans were implemented. Our experimental results showed that cells near the spiral scan center experienced obvious photodamage, whereas cells remained alive over the entire region under the Lissajous beam scanning, confirming the predicted advantage offered by the Lissajous scan over this spiral scan in an endomicroscopy setting.
Journal of Biomedical Optics 02/2012; 17(2):021108. · 3.16 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report an all-fiber-optic scanning, multimodal endomicroscope capable of simultaneous optical coherence tomography (OCT) and two-photon fluorescence (TPF) imaging. Both imaging modalities share the same miniature fiber-optic scanning endomicroscope, which consists of a double-clad fiber with a core operating in single mode at both the OCT (1310 nm) and two-photon excitation (1550 nm) wavelengths, a piezoelectric two-dimensional fiber-optic beam scanner, and a miniature aspherical compound lens suitable for simultaneous acquisition of en face OCT and TPF images. A fiber-optic wavelength division multiplexer was employed in the integrated platform to combine the low coherence OCT light source and the femtosecond two-photon excitation laser into the same optical path. Preliminary imaging results of cell cultures and mouse tissue ex vivo demonstrate the feasibility of simultaneous real-time OCT and TPF imaging in a scanning endomicroscopy setting for the first time.
Optics Letters 02/2012; 37(3):362-4. · 3.40 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Traditionally, charge coupled device (CCD) based image sensors have held sway over the field of biomedical imaging. Complementary metal oxide semiconductor (CMOS) based imagers so far lack sensitivity leading to poor low-light imaging. Certain applications including our work on animal-mountable systems for imaging in awake and unrestrained rodents require the high sensitivity and image quality of CCDs and the low power consumption, flexibility and compactness of CMOS imagers. We present a 132×124 high sensitivity imager array with a 20.1 μm pixel pitch fabricated in a standard 0.5 μ CMOS process. The chip incorporates n-well/p-sub photodiodes, capacitive transimpedance amplifier (CTIA) based in-pixel amplification, pixel scanners and delta differencing circuits. The 5-transistor all-nMOS pixel interfaces with peripheral pMOS transistors for column-parallel CTIA. At 70 fps, the array has a minimum detectable signal of 4 nW/cm(2) at a wavelength of 450 nm while consuming 718 μA from a 3.3 V supply. Peak signal to noise ratio (SNR) was 44 dB at an incident intensity of 1 μW/cm(2). Implementing 4×4 binning allowed the frame rate to be increased to 675 fps. Alternately, sensitivity could be increased to detect about 0.8 nW/cm(2) while maintaining 70 fps. The chip was used to image single cell fluorescence at 28 fps with an average SNR of 32 dB. For comparison, a cooled CCD camera imaged the same cell at 20 fps with an average SNR of 33.2 dB under the same illumination while consuming over a watt.
IEEE Transactions on Biomedical Circuits and Systems 10/2011; 5(5):449-458. · 2.03 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We present a Fourier domain mode locking (FDML) fiber laser with a feedback loop allowing automatic startup without a priori knowledge of the fundamental drive frequency. The feedback can also regulate the drive frequency making the source robust against environmental variations. A control system samples the energy of the light traversing the FDML cavity and uses a voltage controlled oscillator (VCO) to drive the tunable fiber Fabry-Perot filter in order to maximize that energy. We demonstrate a prototype self-starting, self-regulating FDML operating at 40 kHz with a full width tuning range of 140 nm around 1305 nm and a power output of ~40 mW. The laser starts up with no operator intervention in less than 5 seconds and exhibits improved spectral stability over a conventional FDML source. In OCT applications the source achieved over 120 dB detection sensitivity and an ~8.9-µm axial resolution.
Biomedical Optics Express 07/2011; 2(7):2005-11. · 2.33 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report the novel use of laser speckle contrast imaging (LSCI) at multiple exposure times (meLSCI) for enhanced in vivo imaging of the microvascular changes that accompany angiogenesis. LSCI is an optical imaging technique that can monitor blood vessels and the flow therein at a high spatial resolution without requiring the administration of an exogenous contrast agent. LSCI images are obtained under red (632 nm) laser illumination at seven exposure times (1-7 ms) and combined using a curve-fitting approach to obtain high-resolution meLSCI images of the rat brain vasculature. To evaluate enhancement in in vivo imaging performance, meLSCI images are statistically compared to individual LSCI images obtained at a single exposure time. We find that meLSCI reduced the observed variability in the LSCI-based blood-flow estimates by 30% and improved the contrast-to-noise ratio in regions with high microvessel density by 41%. The ability to better distinguish microvessels, makes meLSCI uniquely suited to longitudinal imaging of changes in the vascular microenvironment induced by pathological angiogenesis. We demonstrate this utility of meLSCI by sequentially monitoring, over days, the microvascular changes that accompany wound healing in a mouse ear model.
Journal of Biomedical Optics 05/2011; 16(5):056006. · 3.16 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We present an all-fiber-optic scanning multiphoton endomicroscope with 1.55 μm excitation without the need for prechirping femtosecond pulses before the endomicroscope. The system consists of a 1.55 μm femtosecond fiber laser, a customized double-clad fiber for light delivery and fluorescence collection, and a piezoelectric scan head. We demonstrate two-photon imaging of cultured cells and mouse tissue, both labeled with indocyanine green. Free-space multiphoton imaging with near-IR emission has previously shown benefits in reduced background fluorescence and lower attenuation for the fluorescence emission. For fiber-optic multiphoton imaging there is the additional advantage of using the soliton effect at the telecommunication wavelengths (1.3-1.6 μm) in fibers, permitting dispersion-compensation-free, small-footprint systems. We expect these advantages will help transition multiphoton endomicroscopy to the clinic.
Optics Letters 04/2011; 36(7):1299-301. · 3.40 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Laser speckle contrast imaging (LSCI) has classically been used to image regional blood flow changes in animal models. In this paper, we demonstrate the use of LSCI for elucidating blood flow characteristics in individual microvessels with diameters as small as 24µm. We extracted profiles of speckle contrast values within individual vessels, both along their diameters and along their lengths and inferred that they could be attributed to the flow within the vessel. Profiles along the diameter of vessels revealed maxima at the center of vessels, consistent with fluid dynamics. These observed profiles could be fitted with parabolic curves with a mean coefficient of determination of 0.92. Similarly, analysis of speckle contrast values in the axial direction revealed profiles that progressively decreased in discreet quanta at branch points indicating blood flow bifurcations. Flow estimates obtained from speckle contrast values within branches of vessels obeyed the law of mass conservation with a mean error of only 3.5%. This allowed us to elucidate the percentage distribution of blood flow into each of the downstream branches. This ability of LSCI to resolve blood flow distribution in branching microvessel trees in a minimally invasive and dye free environment over a wide field of view promises to find application in both the neuroscience laboratory as well as intraoperative neurosurgery.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 01/2010; 2010:1978-81.
-
[show abstract]
[hide abstract]
ABSTRACT: Imaging in awake, behaving animals is an emerging field that offers the advantage of being able to study physiological processes and structures in a more natural state than what is possible in tissue slices or even in anesthetized animals. To date, most imaging in awake animals has used optical fiber bundles or electrical cables to transfer signals to traditional imaging-system components. However, the fibers or cables tether the animal and greatly limit the kind and duration of animal behavior that can be studied using imaging methods. We present an integrated imaging microscope (IIM) that incorporates all aspects of an imaging system - illumination, optics and photodetection - into a small footprint device, occupying under 4 cm(3) and weighing 5.4 g, that can be attached to the skull for imaging the brain in mobile rats. Power supply and image storage sufficient for approximately 7 hour operation at 15 frames/s was implemented on a backpack weighing 11.5 g. We implemented several optical techniques including reflectance, spectroscopy, speckle and fluorescence with the IIM, imaged vessels down to 15-20 microm in diameter and obtained, to the best of our knowledge, the worlds first cortical images from an untethered, freely-moving rat.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 01/2010; 2010:5795-8.
-
[show abstract]
[hide abstract]
ABSTRACT: The ability to observe functional and morphological changes in the brain is critical in understanding behavioral and developmental neuroscience. With advances in electronics and miniaturization, electrophysiological recordings from awake, behaving animals has allowed investigators to perform a multitude of behavioral studies by observing changes as an animal is engaged in certain tasks. Imaging offers advantages of observing structure as well as function, and the ability to monitor activity over large areas. However, imaging from an awake, behaving animal has not been explored well. We present the design and characterization of a miniaturized epi-illuminated optical system that is part of a larger goal to perform optical imaging in awake, behaving animals. The system comprises of a tunable light source and imaging optics in a small footprint of 18 mm diameter, 18 mm height and weight 5.7 grams. It offers a spatial illumination non-uniformity of 3.2% over a maximum field of view of 1.5 mm x 1.5 mm, negligible temporal illumination and temperature variation and controllable magnification. Uncorrected radial distortion was 5.3% (corrected to 1.8%) and the spatial frequency response was comparable to a reference system. The system was used to image cortical vasculature in an anesthetized rat.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 01/2009; 2009:5369-72.
-
[show abstract]
[hide abstract]
ABSTRACT: Electrical signals recorded from the brain cover a wide range of amplitudes, frequencies, and spatial scales, from spikes and local field potentials (LFP) inside the brain to electrocorticograms (ECoG) and electroencepalograms (EEG) outside. Each of these signal modalities represent different aspects of neural dynamics that can be combined to infer brain state and function in a broader context.We present a 16-channel interface circuit fabricated in a 0.5 mum CMOS process for the selective acquisition and digitization of any of the modalities. Each channel features a fixed gain bandpass amplifier with a tunable frequency response which allows isolation of the signal of interest without hardware modification and a programmable gain/resolution analog to digital converter (ADC). The bandpass amplifier analog front end has an input referred noise of 1.94 microV(rms) for a bandwidth of 8.2 kHz while drawing 12.2 microA of current from a 3.3 V supply. Experimental recordings with the system show spike signals in rat somatosensory cortex as well as alpha EEG activity in a human subject.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 02/2008; 2008:2741-4.
-
[show abstract]
[hide abstract]
ABSTRACT: Non invasive brain-computer interfaces (BCI) allow people to communicate by modulating features of their electroencephalogram (EEG). Spatiotemporal filtering has a vital role in multi-class, EEG based BCI. In this study, we used a novel combination of principle component analysis, independent component analysis and dipole source localization to design a spatiotemporal multiple source tuning (SPAMSORT) filter bank, each channel of which was tuned to the activity of an underlying dipole source. Changes in the event-related spectral perturbation (ERSP) were measured and used to train a linear support vector machine to classify between four classes of motor imagery tasks (left hand, right hand, foot and tongue) for one subject. ERSP values were significantly (p<0.01) different across tasks and better (p<0.01) than conventional spatial filtering methods (large Laplacian and common average reference). Classification resulted in an average accuracy of 82.5%. This approach could lead to promising BCI applications such as control of a prosthesis with multiple degrees of freedom.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 02/2006; 1:327-30.
-
[show abstract]
[hide abstract]
ABSTRACT: Sensing neurotransmitters is critical in studying neural pathways and neurological disorders. An integrated device is presented which incorporates a potentiostat and a power harvesting and telemetry module. The potentiostat features 16 channels with multiple scales from microamperes to picoamperes. The wireless module is able to harvest power through inductively coupled coils and uses the same link to transmit data to and from the potentiostat. An integrated prototype is fabricated in CMOS technology, and experimentally characterized. Test results show RF powering introduces noise levels of 0.42% and 0.18% on potentiostat current scales of 500pA and 4nA respectively. Real-time multi-channel acquisition of dopamine concentration in vitro is performed with carbon fiber sensors.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 02/2005; 7:7329-32.
-
[show abstract]
[hide abstract]
ABSTRACT: Neurotransmitter sensing is critical in studying nervous pathways and neurological disorders. A 16-channel current-measuring VLSI potentiostat with multiple ranges from picoamperes to microamperes is presented for electrochemical detection of electroactive neurotransmitters like dopamine, nitric oxide etc. The analog-to-digital converter design employs a current-mode, first-order single-bit delta-sigma modulator architecture with a two-stage, digitally reconfigurable oversampling ratio for ranging the conversion scale. An integrated prototype is fabricated in CMOS technology, and experimentally characterized. Real-time multi-channel acquisition of dopamine concentration in vitro is performed with a microfabricated sensor array.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 02/2004; 6:4063-6.
-
[show abstract]
[hide abstract]
ABSTRACT: Spike sorting of neural data from single electrode recordings is a hard problem in machine learning that relies on significant input by human experts. We approach the task of learning to detect and classify spike waveforms in additive noise using two stages of large margin kernel classification and probability regression. Controlled numerical experiments using spike and noise data extracted from neural recordings indicate significant improvements in detection and classification accuracy over linear amplitude- and template-based spike sorting techniques.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 02/2004; 1:546-9.
-
IEEE Engineering in Medicine and Biology Magazine 24(6):23-9. · 2.06 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Previous studies have implicated the abnormal activation of the trigeminal system to be a factor in the pathogenesis of migraine. The relationship between vascular changes and migraine, however, is under considerable debate. In this study, temporal laser speckle contrast imaging is combined with ridge tracking based vessel detection to obtain high resolution (6.7 μm × 6.7 μm), high contrast images of cerebral vascular structure. For the first time, the vasomotor and blood flow responses to electrical stimulation in rat peripheral trigeminal system were obtained simultaneously. The system is capable of picking up individual vessels with diameters down to 30 μm. The spatial spread of the blood velocity response relative to the point of stimulation was studied. Analysis of branching vessels showed a 50 ± 5% vs. 30 ± 5% change in mean peak magnitude and a 54% per second vs. 17% per second change in mean rate of increase for vessels proximal vs. distal to the stimulation site. The penetration depth of the laser used was proven to be sufficient to image dural as well as cortical vessels through a thinned skull preparation. Different responses were observed from cortical and dural vessels. While the diameter of cortical vessels did not change in response to the stimulation the blood velocity went up by 65 ± 5% per second. Dural vessels enlarged by 40 ± 8% and the blood velocity increased by 50 ± 5%. The method described here could be very useful in understanding and studying disorders in the neurovascular system.
Journal of Neuroscience Methods.
