2009 — 2013 |
Morrison, Barclay (co-PI) [⬀] Banta, Scott [⬀] Hillman, Elizabeth |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Directed Evolution of Specific Cell Penetrating Peptides
This award is funded under the American Recovery and Reinvestment Act of 2009(Public Law 111-5).
0853946 Banta
Intellectual merit
The intellectual merit of this proposal results from the first demonstration of the directed evolution of specific cell penetrating peptides (SCPPs). The directed evolution technique offers tremendous potential for the engineering of new peptides, but the critical challenge in this approach is the development of appropriate selection procedures. The PIs will use a novel bi-functional platform based on phage display technology, and this will be coupled with a physiologically relevant selection protocol. Once optimized, this platform will allow one to rapidly evolve valuable new SCPP sequences.
Broader impacts
The broader impacts of this project arise from the new experimental manipulations and therapeutic treatment options that will be made possible by the novel SCPPs. Recent research efforts have produced many exciting mechanism-based therapies. However, the delivery of these therapies to their targets has been a significant limitation. The new SCPPs will enable the expansion of the armamentarium of potential therapies for these devastating diseases. This interdisciplinary research project will also provide a fertile environment for the teaching, training, and mentoring of new engineers both at Columbia University and in the surrounding community.
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1 |
2010 — 2016 |
Hillman, Elizabeth |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Career: Interventional Microscopy For in-Vivo Investigations of Brain Function
0954796 Hillman
The proposed program will develop and refine a platform for in-vivo "Interventional Microscopy". The vision is to create an in-vivo microscope that can not just observe living cells in their natural environment, but directly interact with them. To achieve this they will implement photo-manipulation techniques more commonly applied to in-vitro studies, and translate them for use in-vivo in small animals. By refining experimental paradigms and demonstrating the efficacy of Interventional Microscopy techniques, they will introduce these potentially transformative new tools to a broad audience of biomedical researchers. To achieve these goals, they will start by developing a combined two-photon microscopy and photomanipulation instrument, based around an existing home-built in-vivo two-photon system. The design incorporates two independently focused and shaped beams from two different pulsed lasers, and will be operated by custom-written software. They will then begin developing and optimizing in-vivo techniques for using photoactivable compounds, as well as exploring optical trapping and particle-manipulation techniques in-vivo. To achieve, this they will develop and optimize procedures for in-vivo loading of photomanipulatable substances, as well as developing calibration techniques and controls to ensure that lightactivation in-vivo is reliable and well controlled. They plan to apply these techniques to their ongoing in-vivo studies of neurovascular coupling; the interrelation between blood flow and neuronal activity in the brain. Interventional Microscopy techniques will allow them to selectively manipulate the cells and vessels of the living brain, allowing in depth characterization of fundamental chemical and cellular mechanisms.
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1 |
2011 — 2012 |
Elson, Daniel Hillman, Elizabeth Thompson, Reid |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Conference: Advances in Optics For Biotechnology, Medicine and Surgery Xii, June 5-8, 2011, Naples, Florida
1105324, Hillman
Advances in Optics for Biotechnology, Medicine and Surgery is a small (~120 attendees), interdisciplinary conference focused on the field of biomedical optics. The conference will be held from the 5-8th June 2011 at Naples Beach Hotel, Naples, Florida, USA. Sessions will be composed of invited talks from senior and rising experts focusing on key and emerging aspects of the biomedical optics field. A poster session will provide students and post-docs with ample opportunity to describe and discuss their work, and free-time and social activities will stimulate and promote networking. Substantial effort will be devoted to ensuring geographic, disciplinary, racial, gender and experiential diversity of our participants. Funding from the NSF will allow us to offset the registration costs of 4 students, 4 fellows and young investigators and 8 speakers. Biomedical optics encompasses the interdisciplinary development of new technologies that harness fundamental interactions between light and tissue. The field has already generated a wide range of imaging, diagnostic and therapeutic approaches that have impacted modern medicine. Biomedical optics has also had a profound influence on biomedical research, with advances in microscopy and in-vivo imaging tools keeping pace with the rapid development of transgenic fluorescent-protein in-vivo models and contrast agents with molecular specificity. With these two areas combined, biomedical optics is a continually evolving field impacting both medicine and the basic sciences.
Intellectual Merit: Biomedical optics is a term used to broadly describe almost any technology that exploits light for biomedical applications. The fundamental physics of light transport in tissue and fluorescence and absorption spectroscopy remain at the field's core, yet every year sees the evolution of new technologies for light generation, detection and manipulation as well as new contrast agents, algorithms and biomedical applications, all of which quickly advance the field into new areas. Examples of this in the past 2-3 years include super-resolution microscopy approaches such as PALM and STORM, as well as opto-genetics; a new biological technique that allows excitatory cells to be switched on and off using light.
So while this meeting will be the 12th in a series that has been running for over 20 years, every year has seen continual evolution of the program to encompass cutting edge new advances in the field. We recognize, for example, that it is important for experts in optics to understand the principles, potential, and implications of new approaches such as opto-genetics, since their expertise could significantly improve the way that these valuable new tools can be utilized. Similarly, introducing chemists to the different optical approaches to implementing super-resolution microscopy may lead to improved strategies for developing novel contrast agents. The unique format of this small meeting is highly conducive to intensive training at the cutting edge of our field, allowing new concepts to be introduced and then openly discussed.
Broader Impacts: Biomedical optics advances are generally borne from the juxtaposition of engineering, physics, mathematics, biology, chemistry and clinical medicine. To train in this highly interdisciplinary field, one must master all aspects of optics, device development, data acquisition, modeling and analysis, in addition to a thorough understanding of the biomedical problems being addressed. Many breakthroughs have resulted from simply pairing the right technique with the right biomedical application; realizations that require open interdisciplinary discussion. Widespread adoption of new biomedical optics technologies also requires that end users receive training that allows them to appreciate the fundamental principles governing operation of a particular technology, and therefore both its benefits and limitations.
As such, our field relies upon meetings that draw together experts from all facets of biomedical optics research, including scientists, engineers, clinicians and industry experts, to enable this cross-fertilization. The conference for which we are requesting support serves exactly this purpose: To provide a forum for sharing creative ideas across broad areas of engineering, basic science and medicine, as well as an opportunity for students, trainees and young investigators to diversify their knowledge and develop relationships with key leaders in the field.
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1 |
2011 — 2016 |
Yuste, Rafael (co-PI) [⬀] Hillman, Elizabeth Shepard, Kenneth [⬀] |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Idbr: Cmos Cameras For High-Frame-Rate Time-Correlated Single-Photon Counting
IDBR: CMOS cameras for high-frame-rate time-correlated single-photon counting
Recent advances in biological imaging techniques, particularly those exploring molecular dynamics, are outpacing technological innovation. Fluorescence lifetime holds great potential as a biomarker that can reveal changes in a fluorophore's local chemical and physical environment, as well as the binding dynamics of single proteins through excited state interactions and Förster resonance energy transfer (FRET). Many of the latest active dyes, molecular probes and even transgenic labeling strategies exploit FRET to enable real-time observation of cellular processes both in-vitro and in-vivo. While FRET can be detected using intensity-only measurements, quantitation can be dramatically impaired by experimental factors such as photobleaching, whereas lifetime-based FRET measurements are significantly more robust. Nevertheless, adoption and widespread use of fluorescence lifetime imaging microscopy (FLIM) for biological research has been hindered by two major factors: the speed with which FLIM images can be acquired and the cost and complexity of the instrumentation required for FLIM. In this multidisciplinary proposal, a novel two-dimensional high-frame-rate complementary metal-oxide-semiconductor (CMOS) fluorescent lifetime camera chip based on single-photon avalanche diodes (SPADs) will be developed. This chip will be applied to both wide-field and laser-scanning-based microscopy techniques to enable several important advances in FLIM imaging. In widefield imaging, this will result in acquisition of images at a incident-photon-limited frame rate as high as 1 kHz.
Solid-state imagers are based primarily on two technologies, charged-coupled device (CCD) and CMOS. Both of these imaging technologies are based on converting photons to electrons and collecting many of these electrons to produce a measurable signal. These imagers are now employed in digital cameras of every type, from cell phone cameras to the high-end cameras employed in biological imaging. Since optical techniques are so pervasive in probing biological systems, cameras represent the fundamental interface between the biological world and the solid-state world. In this effort, an entirely new camera chip will be designed based on a device that, instead of collecting electrons produced by photons, counts them, one-by-one. This enables very high sensitivity for photon detection. At the same time it allows resolution of very short (and dim) optical events (on the order of 10's of ps). Such capabilities will enable new types of biological imaging applications. This project supports the multidisciplinary training of graduate and undergraduate students and a significant K-12 outreach effort.
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1 |
2015 |
Rogan, Elizabeth Hillman, Elizabeth |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Optics and the Brain Topical Meeting @ Optical Society of America
PI: Rogan, Elizabeth Proposal No.: 1540895
The high level of technical development stimulated by the BRAIN initiatives is drawing new researchers into the field from diverse disciplines ranging from chemistry and genetics, to laser optics and neurosurgery. OSA has identified the urgent need for a forum where these researchers can come together and discuss optics in the brain at many levels.
The researchers will participate in discussions of highly technical components of optical design and optical interactions with tissue. They will also consider the motivating factors in developing optical tools for neuroscience research across organisms, from cells and flies to rodents and humans. This meeting will provide a forum for presentation of results from projects related to the BRAIN initiative and Human Brain Project, and will stimulate new ideas and collaborations that will provide visionary direction for future projects. In addition this meeting will provide an opportunity for students, trainees and young investigators to present their work, and to hear and network with internationally-renowned invited speakers who represent the broad diversity of this burgeoning field. This meeting will serve to shape the careers of young trainees embarking on careers in which optics and the brain converge. The Optics and the Brain meeting will be held at the Pinnacle Vancouver Harbourside Hotel, Vancouver, Canada April 12-15, 2015.
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0.91 |
2015 — 2017 |
Hillman, Elizabeth M |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Scape Microscopy For High-Speed in-Vivo Volumetric Microscopy in Behaving Organisms @ Columbia University Health Sciences
? DESCRIPTION (provided by applicant): Despite the growing availability of optical markers of neuronal activity, as well as genetic tools for optical manipulation, current optical microscopy techniques for imaging the intact brain at cellular resolution have approached their limits, particularly in terms of 3D volumetric imaging speeds. The brain and nervous system is inherently 3D, with cortical layers playing specific roles in information processing. Small organisms such as Drosophila melanogaster (fruit fly), Danio rerio (zebrafish) and Caenorhabditis elegans, have become valuable platforms for neuroscience research and genetic manipulation, and offer the chance to capture the entire nervous system of a complete, behaving organism. However, for both rodent brain and small organism microscopy, current techniques are limited to slow volumetric imaging rates, or single-plane acquisition. We recently developed a transformative new approach to high speed 3D microscopy called Swept, Confocally-Aligned Planar Excitation (SCAPE) microscopy. SCAPE was conceived as a way to dramatically improve volumetric imaging speeds, while maintaining a simple optical layout and image acquisition geometry. SCAPE is a hybrid between light-sheet microscopy and laser scanning confocal which overcomes the major speed barriers of both techniques. Recently published in Nature Photonics, SCAPE can image at volume rates 10-100 x faster than laser scanning microscopy or fast light-sheet imaging. We have demonstrated imaging of cellular-level structure and function in both the awake, behaving rodent brain and freely moving Drosophila melanogaster larvae at 10-20 volumes per second (VPS) over large fields of view. A further feature of SCAPE is its simple, single, stationary objective, permitting 3D imaging with no motion at the sample, making it well suited for integration with pattered optogenetic manipulation of cells during high-speed 3D imaging. Having achieved `proof of concept' we now wish to develop SCAPE into a tool for routine use by neuroscientists working in both small organisms, for in-toto imaging of cellular activity and behavior, and in awake, behaving mouse brain. The former will be achieved through development and translation of an improved beta prototypes `1P-SCAPE' system, with development of user friendly acquisition software, data handling and analysis platforms, and ultimately its deployment and support for use in studies of somatosensory integration in adult and larval Drosophila. For mouse brain imaging, we propose to test the limits of SCAPE by exploring two- photon implementation (2P-SCAPE), which will afford deeper penetration imaging into scattering tissues such as the rodent brain.
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0.958 |
2016 |
Hillman, Elizabeth M |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Administrative Supplement to Brain Grant U01ns094296 @ Columbia University Health Sciences
? DESCRIPTION (provided by applicant): Despite the growing availability of optical markers of neuronal activity, as well as genetic tools for optical manipulation, current optical microscopy techniques for imaging the intact brain at cellular resolution have approached their limits, particularly in terms of 3D volumetric imaging speeds. The brain and nervous system is inherently 3D, with cortical layers playing specific roles in information processing. Small organisms such as Drosophila melanogaster (fruit fly), Danio rerio (zebrafish) and Caenorhabditis elegans, have become valuable platforms for neuroscience research and genetic manipulation, and offer the chance to capture the entire nervous system of a complete, behaving organism. However, for both rodent brain and small organism microscopy, current techniques are limited to slow volumetric imaging rates, or single-plane acquisition. We recently developed a transformative new approach to high speed 3D microscopy called Swept, Confocally-Aligned Planar Excitation (SCAPE) microscopy. SCAPE was conceived as a way to dramatically improve volumetric imaging speeds, while maintaining a simple optical layout and image acquisition geometry. SCAPE is a hybrid between light-sheet microscopy and laser scanning confocal which overcomes the major speed barriers of both techniques. Recently published in Nature Photonics, SCAPE can image at volume rates 10-100 x faster than laser scanning microscopy or fast light-sheet imaging. We have demonstrated imaging of cellular-level structure and function in both the awake, behaving rodent brain and freely moving Drosophila melanogaster larvae at 10-20 volumes per second (VPS) over large fields of view. A further feature of SCAPE is its simple, single, stationary objective, permitting 3D imaging with no motion at the sample, making it well suited for integration with pattered optogenetic manipulation of cells during high-speed 3D imaging. Having achieved `proof of concept' we now wish to develop SCAPE into a tool for routine use by neuroscientists working in both small organisms, for in-toto imaging of cellular activity and behavior, and in awake, behaving mouse brain. The former will be achieved through development and translation of an improved beta prototypes `1P-SCAPE' system, with development of user friendly acquisition software, data handling and analysis platforms, and ultimately its deployment and support for use in studies of somatosensory integration in adult and larval Drosophila. For mouse brain imaging, we propose to test the limits of SCAPE by exploring two- photon implementation (2P-SCAPE), which will afford deeper penetration imaging into scattering tissues such as the rodent brain.
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0.958 |
2017 — 2021 |
Hillman, Elizabeth M |
U19Activity Code Description: To support a research program of multiple projects directed toward a specific major objective, basic theme or program goal, requiring a broadly based, multidisciplinary and often long-term approach. A cooperative agreement research program generally involves the organized efforts of large groups, members of which are conducting research projects designed to elucidate the various aspects of a specific objective. Substantial Federal programmatic staff involvement is intended to assist investigators during performance of the research activities, as defined in the terms and conditions of award. The investigators have primary authorities and responsibilities to define research objectives and approaches, and to plan, conduct, analyze, and publish results, interpretations and conclusions of their studies. Each research project is usually under the leadership of an established investigator in an area representing his/her special interest and competencies. Each project supported through this mechanism should contribute to or be directly related to the common theme of the total research effort. The award can provide support for certain basic shared resources, including clinical components, which facilitate the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence. |
Characterizing Long-Range Cortical and Subcortical Dynamics in Relation to Corticospinal Output and Motor Control @ Columbia University Health Sciences
Abstract Even a simple movement, like the extension or flexion of a forelimb, requires the activation requires the input of activity from many different neural populations across different brain areas onto motor control centers that control muscle activity. Although many brain areas have been shown to have motor related activity, and to be involved in movement preparation and execution, the relation between activity in these brain areas and the output of the brain onto the spinal cord remain elusive. Therefore, it is absolutely essential to characterize the contribution of upstream neural populations from other cortical and subcortical areas to the activity of the projection-specific populations of corticospinal neurons characterized in Project 1, during different modes of motor control. We propose to perform this characterization using a range of functional imaging approaches that interrogate this problem at different scales. We will utilize wide-field optical mapping (WFOM) to image, in an unbiased manner, the activity hundreds to thousands of cells across many cortical areas in relation to the activity of particular CSNs, which represent the output of the cortex to the spinal cord. Informed by the WFOM imaging, we will then perform simultaneous imaging of populations of neurons, with single cell resolution, in different brain areas using 2-photon random access mesoscopic (2p-RAM) imaging. Finally, we propose to use the 2pRAM mesoscope to simultaneously image the activity of thalamus or striatum through GRIN lenses, and the activity of corticospinal neurons, during particular motor control modes. The knowledge gained in this project will be critical to understand principles governing activity in motor cortex versus other cortical areas in relation to motor output as proposed in project 3, as well as to inform the modelling in project 4. Furthermore, it will help generate predictions that will be tested using closed-loop optogenetic manipulation experiments in project 5.
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0.958 |
2017 |
Hillman, Elizabeth M |
RF1Activity Code Description: To support a discrete, specific, circumscribed project to be performed by the named investigator(s) in an area representing specific interest and competencies based on the mission of the agency, using standard peer review criteria. This is the multi-year funded equivalent of the R01 but can be used also for multi-year funding of other research project grants such as R03, R21 as appropriate. |
Decoding the Neural Basis of Resting-State Functional Connectivity Mapping @ Columbia University Health Sciences
Abstract Resting state functional magnetic resonance imaging (rs-fMRI) is an important modality for imaging the human brain. Capturing fluctuations in the blood oxygen level dependent (BOLD) signal while the brain is `at rest', rs- fMRI can detect distant, and often bilaterally-symmetric regions where activity is synchronized. Such regions are inferred to have `functional connectivity', and patterns of these networks have been found to be altered in a wide range of otherwise indistinguishable disease states. However, despite widespread use of rs-fMRI, interpretation of functional connectivity networks is limited by: 1) The dependence of fMRI BOLD signals on hemodynamic changes as a proxy for neural activity and: 2) A limited understanding of the mechanistic basis of functional connectivity networks in the context of cellular-level interactions and neural representations. In a recently published study, we demonstrated a new optical imaging technique capable of capturing both neural activity and hemodynamics across the bilaterally exposed superficial cortex of awake, behaving mice. This method revealed striking patterns of resting-state neural activity in the awake brain, exhibiting bilateral symmetry and features consistent with resting-state networks. Moreover, we demonstrated that this `neural network activity' was predictive of patterns of resting-state hemodynamics (via a linear convolution model), suggesting that we were visualizing the neural basis of resting state functional connectivity mapping. In the current proposal, we plan to leverage this new view of neural network activity in the brain, to characterize its cellular dependencies, pathways, drivers, behavioral correlates and interactions with hemodynamics. Data will be acquired using novel measurement and circuit manipulation techniques in awake, behaving mice, in addition to analysis of human rs-fMRI, intracranial and intraoperative electrophysiology in patients undergoing epilepsy evaluation (data from ongoing trials) as well as new intraoperative simultaneous optical hemodynamic and electrocorticography recordings. A major aspect of this project will be the aggregation of this data to generate predictive mechanistic and mathematical models of 1) Neural network activity and its dynamic properties and representations across scales and modalities and 2) The coupling relationships between resting-state activity in specific cell types and hemodynamics. These models will be used to derive and test improved methods for rs-fMRI acquisition, analysis and interpretation. To perform this work, we have assembled a world-class interdisciplinary team consisting of neuroscientists, neuroengineers, neurosurgeons, statisticians and experts in resting state fMRI acquisition and analysis. With a sharper understanding of the properties of neural network activity, its dependencies, and how best to harness it in human rs-fMRI, the results of this work could ultimately provide a mechanistic basis for network dysfunctions and their cognitive and behavioral manifestations in disease, potentially yielding new targets for therapies and more robust rs-fMRI based disease detection.
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0.958 |
2019 — 2020 |
Brenner, David Jonathan [⬀] Hillman, Elizabeth M |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Flexible Tools For Pre-Clinical Studies to Answer Key Questions Underlyingheavy-Ion Radiotherapy @ Columbia University Health Sciences
SUMMARY / ABSTRACT Heavy-ion radiation therapy (HIRT) differs from other radiotherapy modalities such as x rays and protons as these high-LET (Linear Energy Transfer) radiations deposit energy far more densely on a microscopic scale. There is currently strong interest in the introduction of HIRT to the U.S., largely based on the experience of carbon-ion radiotherapy in Japan and Germany, where very encouraging survival rates have been reported for a number of hard-to-treat cancers such as pancreas, rectum and sarcomas. For example, 2-year survival of 50 to 65% has been reported after combined carbon-ion and gemcitabine chemotherapy for locally-advanced pancreatic cancer, remarkably encouraging at a post-treatment time when survival is dominated by distant metastases. Thus there has been much discussion that, as well as producing local effects to the tumor, HIRT may also be inducing long-range systemic anti-cancer effects. However, the underlying mechanisms for such high-LET-induced long-range systemic effects are not understood and there is evidence that the classic radiobiological phenomena underlying the efficacy of conventional x-ray radiotherapy, while still potentially relevant for local tumor control, are not the dominant phenomena driving the potential systemic efficacy of HIRT. Rather the data suggest different high-LET-induced mechanisms underlying radiation-induced long- range anti-cancer effects ? and what is not known is the LET dependence of these long-range effects. In this BRP, and leveraging from the unique technologies and skillsets at the Radiological Research Accelerator Facility (RARAF) and the Laboratory for Functional Optical Imaging (LFOI), novel tools will be developed to study and understand long-range radiation-induced biological effects, and particularly their dependence on LET. The key tools will be 1) a series of mono-LET ion beams providing spatially defined 3-D exposures, integrated with 2) SCAPE (Swept Confocally-Aligned Planar Excitation) wide-area 3D microscopy, imaging within and outside the radiation field. In parallel, the BRP tools will be applied to address the central hypothesis of LET dependence of long-range radiation effects. These studies will encompass increasing levels of complexity from tumor cells through in-vitro tumor/tissue models to in-vivo tumor models. To develop and apply these technologies, an interdisciplinary team has been assembled of accelerator physicists and radiobiologists from RARAF, and biomedical engineers from LFOI, enhanced through continuous engagement with internationally recognized scientists and clinicians with experience in HIRT. Apart from the primary goal of optimizing HIRT efficacy, understanding the relevant LET dependencies in HIRT will provide a pathway for determining the optimal ion / ions for its use ? a key outcome that in turn will likely determine the future worldwide usage of HIRT, in that the capital cost of HIRT is dominated by the choice of ion or ions to be used. If, for example, the optimal LET range for HIRT could be achieved with helium ions, a helium therapy machine would be far smaller and cheaper than a >$150M carbon-ion machine.
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0.958 |