Available Innovations
Technology:
This invention combines interferometry and photoactivated localization microscopy (PALM) to create a new light microscopy technique that provides sub 20-nm, three-dimensional localization. PALM is a widely used, super resolution biological imaging technique that improves the spatial resolution of optical microscopy by at least an order of magnitude and is based on imaging and localizing a large number of single fluorescent molecules sequentially in time and reconstructing a high resolution image based on the molecular coordinates. PALM generates two-dimensional images; obtaining 3D PALM images requires another measurement sensitive to the third dimension. The most sensitive method is to perform multiphase interferometery on the fluorescent light of each molecule and thereby extract its position in the third dimension. The technology described here called iPALM for “interferometric” PALM extends the use of PALM bridging the gap between light and electron microscopy.
The 3D super resolution of this technology has been demonstrated on fixed cells, where it imaged cellular ultrastructure seen previously only with electron microscopy. Using this method it was also possible to resolve the 25-nm diameter of microtubules, resolve dorsal and ventral plasma membranes, and visualize the arrangement of integrin receptors in the endoplasmic reticulum. The nanoscale architecture of various proteins in cell adhesion structure has also been measured. This technology employs an optical system with multiple detectors and a processor. As with 2D PALM, switchable optical labels are used such as the photo activatable fluorescent proteins or switchable dyes.
Advantages:
- Opens a third dimension to photoactivated localization microscopy
- Enables observation of molecules at the same resolution as electron microscopy
- Best vertical resolution for given number of photons
- Proven with photoactivatable fluorescent proteins
Applications:
- Resolving protein organization within molecular assemblies in cells
- Deciphering molecular scale architecture and interactions that constitute biological structures
- 3D protein trafficking and diffusion in cytosol
- Correlative microscopy
Publications:
Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure; Shtengel G, Galbraith JA, Galbraith CG, Lippincott-Schwartz J, Gillette JM, Manley S, Sougrat R, Waterman CM, Kanchanawong P, Davidson MW, Fetter RD, Hess HF.; Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3125-30.
Nanoscale architecture of integrin-based cell adhesions; Kanchanawong P, Shtengel G, Parapera A, Ramko E, Davidson M, Hess H, Waterman C; Nature 2010 Nov 25 Vol 468 p 580.
Patent Status: Issued US patent 7,924,432
Desired Partnership: License
Contact and further information: techtransfer@janelia.hhm.org
Technology:
Pulsed lasers are important tools for nonlinear bioimaging and are widely used in techniques such as two-photon fluorescence excitation (TPE) microscopy. A limitation of using pulsed lasers is the tendency for photoinduced damage of samples. The invention described here is a pulse splitter that reduces photobleaching and photodamage in TPE. The pulse splitter receives individual laser pulses and creates a finite number of equal intensity pulses from each pulse input. It is simple, compact and requires little or no adjustment. The splitter can be configured to provide a varied number of pulses (4 to 128).
The value of the pulse splitter has been demonstrated in a variety of samples, both living and fixed. In GFP labeled fixed brain samples the pulse splitter reduced the rate of photobleaching by over fourfold. During in vivo imaging of C. elegans the photobleaching rate with 64x pulse splitting decreased 9-fold. During calcium imaging of rat hippocampal brain slices a six- to 20-fold decrease in the rate of photodamage was observed with a concomitant 6-fold gain in lifetime. This technology is a useful tool for extending the capabilities of TPE and potentially other non-linear imaging techniques.
Applications:
- Imaging of fixed and living samples.
- Calcium imaging
- Multiphoton imaging, including TPE, second harmonic generation, sum frequency generation, CARS and single-molecule TPE microscopy
Advantages:
- Compact
- Requires few adjustments
- Can be flexibly reconfigured to optimize the repetition rate, pulse spacing and pulse intensity
- Adaptable to the large installed base of preexisting lasers and TPE microscopes.
Publication:
High-speed, low-photodamage nonlinear imaging using passive pulse splitters; Ji N, Magee JC, Betzig E.; Nat Methods. 2008 Feb; 5(2): 197-202.
Desired Partnership: License agreement for manufacturing and distribution.
Contact and further information: techtransfer@janelia.hhm.org
This technology combines small molecules and engineered ligand gated ion channels to serve as chemical/genetic tools for controlling electrical activity of molecularly defined neuron populations. It has potential applications as a research tool and as a therapeutic strategy.
Technology:
This technology advances the resolution and depth limit of light microscopy using an innovative and effective approach: adaptive optics (AO). Depth and resolution in life science imaging are limited by the inherent heterogeneity of biological specimens. These specimens give rise to optical distortions, known as aberrations that lead to the loss of signal, image fidelity and resolution. AO has been used in telescopes to address the problem of optical aberration of light beams as they propagate through our atmosphere, but it has been applied to microscopy in only a limited manner due to technical challenges. This new technology is an image based AO method that is simpler to implement than other AO methods and suitable for microscopy. This innovation comprises the use of an SLM in the optical path of a microscope and proprietary algorithms. It attains near diffraction-limited performance from a variety of biological samples with only minor modification of a light microscope.
The value of the technology has been demonstrated in two-photon microcopy, where it attained near-diffraction–limited performance in varied biological and non-biological samples exhibiting aberrations large or small and smoothly varying or abruptly changing. Work in fixed mouse cortical slices illustrates the capability to improve signal and resolution to depths of 500m. The technology is applicable to other imaging modalities, including confocal microscopy and widefield microscopy.
Advantages:
- Improved signal and resolution to depths of 500m
- Near diffraction limited resolution
- Requires only minor modification of the microscope
- Can be used with two photon, confocal and wide field microscopy
Applications:
- Basic life science research
- Imaging of living or thick biological specimens
- In vivo calcium imaging
- Two photon microscopy
- CARS, second harmonic generation and third harmonic generation imaging
Publication:
Ji, N., Milkie, D. E., & Betzig, E., Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues. Nature Methods, Vol. 7 No.2, 141-147 (2010)
Desired Partnership: License
Contact and further information: techtransfer@janelia.hhm.org
Use of a Bessel beam to illuminate a sample with light sheets that are sufficiently thin to achieve isotropic 3D resolution high-speed 3D fluorescent imaging. The technology can visualize cellular processes in spatiotemporal detail not attainable with other fluorescent microscopy techniques.
Technology:
Initial screening of candidate agonists or antagonists for therapeutic protein targets (e.g. GPCRs) is routinely performed by fluorescence imaging using calcium signaling dyes. The invention described here is a multi-color, genetically encoded calcium reporter system with significant advantages over commercially available screening reagents and kits. This technology platform encompasses a series of novel engineered calcium indicators with a broad spectral range (blue, cyan, green, yellow and red). Because they are genetically encoded, they can be integrated into a cell line of interest and targeted to specific cell types (both cultured and primary cells) and sub-cellular locations with minimal phenotypic effects. They provide a high-throughput drug screening format with signal-to-noise ratio (SNR) superior to commercial dyes and kits as demonstrated through systematic comparison with the state-of-the-art small molecule screens. The expression level in integrated cell lines can be stably maintained, minimizing assay variability over well-to-well differences in dye incubation. No expensive dyes or cofactors are required.
The use of these indicators has been demonstrated with the following protein targets: H1R, mGluR5, V2R, 5HT-2C, GnRHR and P2Y receptors, with a battery of common agonists, antagonists, and marketed drugs. The sensors possess sufficient SNR to permit imaging the activation of endogenous protein targets, rather than requiring over-expression, as demonstrated with muscarinic acetylcholine receptors, glutamate receptors and purinergic receptors. The high SNR of the sensors allows imaging activation of GPCRs mediated through G-proteins other than the canonical Gq pathway.
Simultaneous two-color imaging has been demonstrated with red & green sensors, both (1) within the same cell, in different sub-cellular compartments (e.g. nucleus, membrane, pre- and post-synapse), and (2) in different cell types (e.g. primary neurons in red & primary glia in green, and vice versa). These capabilities open up new avenues for drug screening: compounds can now be tested for their interplay between extra-cellular receptor activation and intra-cellular effects. This is not possible with small molecule dyes, as the two signals are convolved. In addition, this technology can also be used for calcium imaging in living cells and organisms, facilitating the identification of potential therapeutic targets in diseases associated with altered calcium homeostasis.
This invention, and cell lines derived from it, will dramatically reduce the costs of initial drug panning experiments. This technology can be used for high-throughput screening for agonists and antagonists to a wide array of additional molecular targets and enable new kinds of assays that were not possible previously, in a wide array of scientific settings. For instance, with targetable, high-SNR, multi-color indicators, it should be feasible to simultaneously image multiple components of stem cell differentiation, set up a viral fusion inhibitor screen, and create very sensitive animal models for the study of neural regeneration, neurodegenerative diseases, infectious diseases and cancer.
Advantages:
- Brighter, more photostable, and greater dynamic range than prior GECIs and currently marketed HTS reagents.
- Better signal to noise, simpler to perform, lower assay-to-assay variability and increased sensitivity compared to dye based kits
- Ability to perform targeted multi-color imaging of different cell types & sub-cellular compartments
- Does not require dyes or cofactors
- Quantitative, linear response
Applications:
- In vivo calcium imaging to monitor neural activity in live cells, tissues and animals
- High throughput screening for agonists and antagonists of therapeutic targets, such as GPCRs, ion channels and endogenous receptors
- Multi-color imaging
- Sub-cellular targeting
Patent Status: Patent application filed
Desired Partnership: License
Contact and further information: techtransfer@janelia.hhm.org
Method for creating biosensors for diverse range of analytes, including some that cannot be easily detected by antibodies or nucleic acids. The affinity for analyte and the fluorescent readout can be independently optimized.
16 channel independent single electrode micro drive for mice that weighs under 4 grams and is stable for free behavior single cell recording. Low profile and ergonomically comfortable for an un-tethered animal.
Technology:
RIVETS is a unique sample preparation and handling system than enables the combining of in vitro and in vivo physiology experiments on one microscope. RIVETS has been designed by scientists and engineers at HHMI’s Janelia Farm Research Campus to address the needs of cutting edge neurophysiology research. The RIVETS system is comprised the following components:
- Metal platform that accommodates both head posts and slice chambers and can be integrated into a microscope stage or independent from the microscope;
- Head posts with removable target system, and slice chamber, that fit interchangeably in the above described platform, and that can be produced cheaply and quickly by the end user on a 3D printer using stl file format;
- Stl design files covering the slice chamber and several head post configurations for different applications;
- The slice chambers and head posts are interchangeable within the same platform, enabling rapid switching between in vitro and in vivo experiments.
Advantages:
- Low cost compared to systems that utilize titanium head posts
- Highly customizable
- Long term stability and biocompatibility
- Rapid conversion between preparations
Applications:
- In vivo and in vitro imaging
- Electrophysiology
- Free moving behavior experiments
Desired Partnership: Manufacturing and distribution agreement
Contact and further information: techtransfer@janelia.hhm.org
Technology:
This technology, the first of its kind, is a 128 channel head stage suitable for use in free moving mouse. This head stage has been designed by scientists and engineers at HHMI’s Janelia Farm Research Campus to address the needs of cutting edge neurophysiology research. This technology will enable neurophysiology experiments in mouse that have heretofore been difficult or impossible to perform at these high channel counts.
The head stage is based on INTAN analog multiplexer chips, acquisition software, and a flexible circuit board and has a total weight of only 1.25-2.21 grams depending on choice of probe input connectors.
This head stage can be used with any currently available probe and has been designed to accommodate new generations of probes under development that will offer increased densities.
Advantages:
- Extremely light weight and compact
- Fully implemented solution
- 8-12 conductor tether cable
- Compatible with all current commercial probes
Applications:
- Standard extracellular recording in awake, freely moving mice
- Electrophysiology imaging
Desired Partnership: Manufacturing and distribution agreement
Contact and further information: techtransfer@janelia.hhm.org
