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University of California, Berkeley

35 ARTICLES PUBLISHED IN JoVE

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Biology

Mechanical Stimulation of Stem Cells Using Cyclic Uniaxial Strain
Kyle Kurpinski 1, Song Li 1
1Department of Bioengineering, University of California, Berkeley

It is widely understood that mechanical forces in the body can influence cell differentiation and proliferation. Here we present a video protocol demonstrating the use of a custom-built bioreactor for delivering uniaxial cyclic tensile strain to stem cells cultured on flexible micropatterned substrates.

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Biology

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy
Ena Ladi 1, Paul Herzmark 1, Ellen Robey 1
1Department of Molecular and Cell Biology, University of California, Berkeley

We present step-by-step instructions for the generation of neonatal chimeras as well as the dissection and preparation of the thymus for ex vivo imaging by 2-Photon Microscopy.

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Biology

Neurocircuit Assays for Seizures in Epilepsy Mutants of Drosophila
Iris C. Howlett 1, Mark A. Tanouye 1,2
1Department of Molecular and Cell Biology, University of California, Berkeley, 2Department of Environmental Science, Policy Management, University of California, Berkeley

Using high frequency electrical stimulation, seizure-like activity can be induced in Drosophila. This activity is easily recorded from the giant fiber system.

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JoVE Journal

Comprehensive Compositional Analysis of Plant Cell Walls (Lignocellulosic biomass) Part I: Lignin
Cliff E. Foster 1, Tina M. Martin 1, Markus Pauly 2
1Great Lakes Bioenergy Research Center, Michigan State University (MSU), 2Great Lakes Bioenergy Research Center and DOE-Plant Research Lab, Michigan State University (MSU)

Plant biomass is a major carbon-neutral renewable resource that could be used for the production of biofuels. Plant biomass consists mainly of cell walls, a structurally complex composite material termed lignocellulosics. Here we describe a protocol for a comprehensive analysis of the content and composition of the polyphenolic lignin.

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JoVE Journal

Comprehensive Compositional Analysis of Plant Cell Walls (Lignocellulosic biomass) Part II: Carbohydrates
Cliff E. Foster 1, Tina M. Martin 1, Markus Pauly 2
1Great Lakes Bioenergy Research Center, Michigan State University (MSU), 2Great Lakes Bioenergy Research Center and DOE-Plant Research Lab, Michigan State University (MSU)

Plant biomass is a major carbon-neutral renewable resource that could be used for the production of biofuels. Plant biomass consists mainly of cell walls, a structurally complex composite material termed lignocellulosics. Here we describe a protocol for a comprehensive analysis of the content and composition of wall derived carbohydrates.

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Biology

OLIgo Mass Profiling (OLIMP) of Extracellular Polysaccharides
Markus Günl 1, Sascha Gille 1, Markus Pauly 1,2
1Energy Biosciences Institute, University of California, Berkeley, 2Department of Plant and Microbial Biology, University of California, Berkeley

A rapid way is described to gain insights into the structure of polysaccharides in an extracellular matrix. The method takes advantage of the specificity of glycosylhydrolases and the sensitivity of mass spectrometry allowing minute amounts of materials to be analyzed. This technique is adaptable to be used directly on tissue itself.

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Biology

Label-free in situ Imaging of Lignification in Plant Cell Walls
Martin Schmidt 1, Pradeep Perera 1, Adam M. Schwartzberg 2, Paul D. Adams 3, P. James Schuck 2
1Energy Biosciences Institute, University of California, Berkeley, 2Molecular Foundry, Lawrence Berkeley National Laboratory, 3Physical Biosciences Division, Lawrence Berkeley National Laboratory

A method based on confocal Raman microscopy is presented that affords label-free visualization of lignin in plant cell walls and comparison of lignification in different tissues, samples or species.

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Immunology and Infection

Automated Separation of C. elegans Variably Colonized by a Bacterial Pathogen
Kwame Twumasi-Boateng 1, Maureen Berg 1, Michael Shapira 1
1Department of Integrative Biology, University of California, Berkeley

The wormsorter facilitates genetic screens in Caenorhabditis elegans by sorting worms according to expression of fluorescent reporters. Here, we describe a new usage: sorting according to colonization by a GFP-expressing pathogen, and we employ it to examine the poorly understood role of pathogen recognition in initiating immune responses.

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Bioengineering

Lipid Bilayer Vesicle Generation Using Microfluidic Jetting
Christopher W. Coyne 1, Karan Patel 1, Johanna Heureaux 1, Jeanne Stachowiak 3, Daniel A. Fletcher 4,5, Allen P. Liu 1,2
1Department of Mechanical Engineering, University of Michigan, 2Department of Biomedical Engineering, University of Michigan, 3Department of Biomedical Engineering, Institute for Cellular and Molecular Biology, The University of Texas at Austin, 4Department of Bioengineering, University of California, Berkeley, 5Physical Biosciences Division, Lawrence Berkeley National Laboratory

Microfluidic jetting against a droplet interface lipid bilayer provides a reliable way to generate vesicles with control over membrane asymmetry, incorporation of transmembrane proteins, and encapsulation of material. This technique can be applied to study a variety of biological systems where compartmentalized biomolecules are desired.

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Bioengineering

Production and Targeting of Monovalent Quantum Dots
Daeha Seo *1,2,3, Justin Farlow *4,5,6, Kade Southard 1,4,7, Young-wook Jun 1,7, Zev J. Gartner 4,5,6,7
1Department of Otolaryngology, University of California, San Francisco, 2Department of Chemistry, University of California, Berkeley, 3Materials Science Division, Lawrence Berkeley National Laboratory, 4Department of Pharmaceutical Chemistry, University of California, San Francisco, 5Tetrad Graduate Program, University of California, San Francisco, 6Center for Systems and Synthetic Biology, University of California, San Francisco, 7Chemistry and Chemical Biology Graduate Program, University of California, San Francisco

We provide detailed instructions for the preparation of monovalent targeted quantum dots (mQDs) from phosphorothioate DNA of defined length. DNA wrapping occurs in high yield, and therefore, products do not require purification. We demonstrate the use of the SNAP tag to target mQDs to cell-surface receptors for live-cell imaging applications.

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Developmental Biology

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
John D. Blair 1, Helen S. Bateup 1, Dirk F. Hockemeyer 1
1Department of Molecular and Cell Biology, University of California, Berkeley

Genome editing of human pluripotent stem cells (hPSCs) can be done quickly and efficiently. Presented here is a robust experimental procedure to genetically engineer hPSCs as exemplified by editing the AAVS1 safe harbor locus to express EGFP and introduce antibiotic resistance.

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Developmental Biology

Microinjection for Transgenesis and Genome Editing in Threespine Sticklebacks
Priscilla A. Erickson 1, Nicholas A. Ellis 1, Craig T. Miller 1
1Department of Molecular and Cell Biology, University of California, Berkeley

Transgenic manipulations and genome editing are critical for functionally testing the roles of genes and cis-regulatory elements. Here a detailed microinjection protocol for the generation of genomic modifications (including Tol2-mediated fluorescent reporter transgene constructs, TALENs, and CRISPRs) is presented for the emergent model fish, the threespine stickleback.

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Biology

Dissection and Flat-mounting of the Threespine Stickleback Branchial Skeleton
Nicholas A. Ellis 1, Craig T. Miller 1
1Department of Molecular and Cell Biology, University of California, Berkeley

The branchial skeleton, including gill rakers, pharyngeal teeth, and branchial bones, serves as the primary site of food processing in most fish. Here we describe a protocol to dissect and flat-mount this internal skeleton in threespine sticklebacks. This method is also applicable to a variety of other fish species.

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Biology

Visualizing Stromule Frequency with Fluorescence Microscopy
Jacob O. Brunkard 1, Anne M. Runkel 2, Patricia Zambryski 2
1Plant Gene Expression Center, Agricultural Research Service, USDA, 2Department of Plant and Microbial Biology, University of California, Berkeley

Protocols to investigate the dynamics of chloroplast stromules, the stroma-filled tubules that extend from the surface of chloroplasts, are described.

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Chemistry

Combining Solid-state and Solution-based Techniques: Synthesis and Reactivity of Chalcogenidoplumbates(II or IV)
Günther Thiele 1, Carsten Donsbach 2, Isabell Nußbruch 2, Stefanie Dehnen 2
1Department of Chemistry, University of California, Berkeley, 2Fachbereich Chemie, Philipps-Universität Marburg and Wissenschaftliches Zentrum für Materialwissenschaften

The synthesis of chalcogenidoplumbates(II,IV) via the in situ reduction of nominal "PbCh2" (Ch = Chalcogen) and via a solid-state reaction and subsequent solvothermal reactions is presented. Additionally, reactivities of plumbate(II) solutions are portrayed, which yield the heaviest-known CO homolog known to date: the µ-PbSe ligand.

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Developmental Biology

A Modified Trier Social Stress Test for Vulnerable Mexican American Adolescents
Megan M. Johnson 1, Julianna Deardorff 1, Kimberly Parra 1, Abbey Alkon 2, Brenda Eskenazi 1, Elizabeth Shirtcliff 3
1Center for Environmental Research and Children's Health (CERCH), Berkley School of Public Health, University of California, Berkeley, 2San Francisco (UCSF) School of Nursing, University of California, San Francisco, 3Human Development and Family Studies, Iowa State University

Here, we present a protocol that provoked cortisol reactivity in a vulnerable adolescent Mexican American sample utilizing a modified version of the Trier Social Stress Test (TSST). Saliva samples were collected at baseline, 15, 30, and 45 min post-TSST onset. Future research could utilize this modified TSST with vulnerable youth.

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Chemistry

Low Pressure Vapor-assisted Solution Process for Tunable Band Gap Pinhole-free Methylammonium Lead Halide Perovskite Films
Carolin M. Sutter-Fella *1,2,3, Yanbo Li *1,4, Nicola Cefarin 1,5,6, Aya Buckley 1,7, Quynh Phuong Ngo 8,9, Ali Javey 2,3, Ian D. Sharp 1, Francesca M. Toma 1
1Joint Center for Artificial Photosynthesis, Chemical Sciences Division, Lawrence Berkeley National Laboratory, 2Electrical Engineering and Computer Sciences, University of California, Berkeley, 3Materials Science Division, Lawrence Berkeley National Laboratory, 4Institute of Fundamental and Frontier Sciences, University of Electronic Science and Technology of China, 5Department of Physics, Graduate School of Nanotechnology, University of Trieste, 6TASC Laboratory, IOM-CNR - Istituto Officina dei Materiali, 7Department of Chemistry, University of California, Berkeley, 8Materials Science and Engineering, University of California, Berkeley, 9Joint Center for Artificial Photosynthesis, Lawrence Berkeley National Laboratory

Here, we present a protocol for the synthesis of CH3NH3I and CH3NH3Br precursors and the subsequent formation of pinhole-free, continuous CH3NH3PbI3-xBrx thin films for the application in high efficiency solar cells and other optoelectronic devices.

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Biology

ODELAY: A Large-scale Method for Multi-parameter Quantification of Yeast Growth
Thurston Herricks 1, Fred D. Mast 1,2, Song Li 1, John D. Aitchison 1,2
1Institute for Systems Biology, 2Center for Infectious Disease Research

We present a method for quantifying growth phenotypes of individual yeast cells as they grow into colonies on solid media using time-lapse microscopy termed, One-cell Doubling Evaluation of Living Arrays of Yeast (ODELAY). Population heterogeneity of genetically identical cells growing into colonies can be directly observed and quantified.

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Environment

Wastewater Irrigation Impacts on Soil Hydraulic Conductivity: Coupled Field Sampling and Laboratory Determination of Saturated Hydraulic Conductivity
Jack E. Watson 1, Tyson Robb 2, Danielle Andrews-Brown 3, Melissa Miller 1
1Department of Ecosystem Science and Management, Pennsylvania State University, 2Department of Geography and Environmental Sustainability, State University of New York, Oneonta, 3Department of Geology and Environmental Science, University of Pittsburgh

Here we present a methodology which matches a soil sample size and a hydraulic conductivity measurement device to prevent the so-called wall flow along the inside of the soil container from being erroneously included in water flow measurements. Its use is demonstrated with samples collected from a wastewater irrigation site.

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Genetics

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Behnom Farboud *1,2, Erin Jarvis *1, Theodore L. Roth *3,4,5,6, Jiyung Shin *1,3, Jacob E. Corn 1,3, Alexander Marson 3,5,6,7,8,9, Barbara J. Meyer 1,2, Nipam H. Patel 1,10, Megan L. Hochstrasser 3
1Department of Molecular Cell Biology, University of California, Berkeley, 2Howard Hughes Medical Institute, University of California, Berkeley, 3Innovative Genomics Institute, University of California, Berkeley, 4Biomedical Sciences Graduate Program, University of California, San Francisco, 5Department of Microbiology and Immunology, University of California, San Francisco, 6Diabetes Center, University of California, San Francisco, 7Chan Zuckerberg Biohub, 8Department of Medicine, University of California, San Francisco, 9UCSF Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, 10Department of Integrative Biology, University of California, Berkeley

Utilizing a preassembled Cas9 ribonucleoprotein complex (RNP) is a powerful method for precise, efficient genome editing. Here, we highlight its utility across a broad range of cells and organisms, including primary human cells and both classic and emerging model organisms.

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JoVE Journal

Exploring the Root Microbiome: Extracting Bacterial Community Data from the Soil, Rhizosphere, and Root Endosphere
Tuesday Simmons 1, Daniel F. Caddell 1, Siwen Deng 1, Devin Coleman-Derr 1,2
1Department of Plant and Microbial Biology, University of California, Berkeley, 2Plant Gene Expression Center, USDA ARS

Here, we describe a protocol to obtain amplicon sequence data of soil, rhizosphere, and root endosphere microbiomes. This information can be used to investigate the composition and diversity of plant-associated microbial communities, and is suitable for the use with a wide range of plant species.

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Genetics

Single Molecule Fluorescence In Situ Hybridization (smFISH) Analysis in Budding Yeast Vegetative Growth and Meiosis
Jingxun Chen 1, David McSwiggen 2, Elçin Ünal 1
1Department of Molecular and Cell Biology, Barker Hall, University of California, Berkeley, 2Department of Molecular and Cell Biology, Li Ka Shing Center, University of California, Berkeley

This single molecule fluorescence in situ hybridization protocol is optimized to quantify the number of RNA molecules in budding yeast during vegetative growth and meiosis.

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Bioengineering

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay
Jana A. Hassan 1, Ilea J. Chau-Ly 1, Jennifer D. Lewis 1,2
1Department of Plant and Microbial Biology, University of California, Berkeley, 2Plant Gene Expression Center, United States Department of Agriculture

The seedling flood assay facilitates rapid screening of wild tomato accessions for the resistance to the Pseudomonas syringae bacterium. This assay, used in conjunction with the seedling bacterial growth assay, can assist in further characterizing the underlying resistance to the bacterium, and can be used to screen mapping populations to determine the genetic basis of resistance.

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Biology

Measurements of Physiological Stress Responses in C. Elegans
Raz Bar-Ziv *1, Ashley E. Frakes *1, Ryo Higuchi-Sanabria *1, Theodore Bolas 1, Phillip A. Frankino 1, Holly K. Gildea 1, Melissa G. Metcalf 1, Andrew Dillin 1
1Department of Molecular and Cell Biology, University of California, Berkeley

Here, we characterize cellular proteotoxic stress responses in the nematode C. elegans by measuring the activation of fluorescent transcriptional reporters and assaying sensitivity to physiological stress.

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Biochemistry

Isolation of Histone from Sorghum Leaf Tissue for Top Down Mass Spectrometry Profiling of Potential Epigenetic Markers
Mowei Zhou 1, Shadan H. Abdali 1, David Dilworth 2, Lifeng Liu 2, Benjamin Cole 2, Neha Malhan 1, Amir H. Ahkami 1, Tanya E. Winkler 1, Joy Hollingsworth 3, Julie Sievert 3, Jeff Dahlberg 3, Robert Hutmacher 4,5, Mary Madera 6, Judith A. Owiti 6, Kim K. Hixson 1, Peggy G. Lemaux 6, Christer Jansson 1, Ljiljana Paša-Tolić 1
1Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, 2DOE-Joint Genome Institute, Lawrence Berkeley Laboratory, 3Kearney Agricultural Research and Extension Center, University of California Agriculture and Natural Resources, 4West Side Research and Extension Center, University of California, 5Department of Plant Sciences, University of California, Davis, 6Department of Plant and Microbial Biology, University of California, Berkeley

The protocol has been developed to effectively extract intact histones from sorghum leaf materials for profiling of histone post-translational modifications that can serve as potential epigenetic markers to aid engineering drought resistant crops.

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Biology

Recording Electrical Currents across the Plasma Membrane of Mammalian Sperm Cells
Boheng Liu 1,2, Nadine Mundt 3,4, Melissa Miller 1, David E. Clapham 5, Yuriy Kirichok 6, Polina V. Lishko 1
1Department of Molecular and Cell Biology, University of California, Berkeley, 2Department of Neurobiology, Peking University, 3Institute for Biology II / Chemosensation Lab, RWTH Aachen University, 4Research Training Group 2416 MultiSenses-MultiScales, RWTH Aachen University, 5Howard Hughes Medical Institute, Janelia Research Campus, 6Department of Physiology, University of California, San Francisco

This protocol describes how to perform electrical recordings from mammalian sperm cells in a whole-cell configuration, with the goal of directly recording ion channel activity. The method has been instrumental in describing the electrophysiological profiles of several sperm ion channels and helped to reveal their molecular identity and regulation.

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Genetics

Surveying Low-Cost Methods to Measure Lifespan and Healthspan in Caenorhabditis elegans
Toni Castro Torres *1, Darius Moaddeli *1, Maxim Averbukh 1, Aeowynn J. Coakley 1, Naibedya Dutta 1, Gilberto Garcia 1, Ryo Higuchi-Sanabria 1
1Leonard Davis School of Gerontology, University of Southern California

Caenorhabditis elegans serve as an excellent model system with robust and low-cost methods for surveying healthspan, lifespan, and resilience to stress.

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Environment

Visualizing Field Data Collection Procedures of Exposure and Biomarker Assessments for the Household Air Pollution Intervention Network Trial in India
Karthikeyan D. Rajamani 1, Sankar Sambandam 1, Krishnendu Mukhopadhyay 1, Naveen Puttaswamy 1, Gurusamy Thangavel 1, Durairaj Natesan 1, Rengaraj Ramasamy 1, Saritha Sendhil 1, Amudha Natarajan 1, Vigneswari Aravindalochan 1, Ajay Pillarisetti 2, Michael Johnson 3, Joshua Rosenthal *4, Kyle Steenland 5, Ricardo Piedhrahita 3, Jennifer Peel 6, Maggie L. Clark 6, Dana Boyd Barr 5, Sarah Rajkumar 6, Bonnie Young 6, Shirin Jabbarzadeh 7, Ghislaine Rosa 8, Miles Kirby 9, Lindsay J. Underhill 10, Anaite Diaz-Artiga 11, Amy Lovvorn 5, William Checkley 12, Thomas Clasen 5, Kalpana Balakrishnan 1
1Department of Environmental Health Engineering, ICMR Center for Advanced Research on Air Quality, Climate and Health, Faculty of Public Health, Sri Ramachandra Institute of Higher Education and Research (Deemed University), 2Division of Environmental Health Sciences, University of California, Berkeley, 3Berkeley Air Monitoring Group, 4Division of International Epidemiology and Population Studies, National Institutes of Health, 5Gangarosa Department of Environmental Health, Rollins School of Public Health, Emory University, 6Department of Environmental and Radiological Health Sciences, Colorado State University, 7Department of Biostatistics and Informatics, Rollins School of Public Health, Emory University, 8Department of Disease Control, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, 9Department of Global Health & Population, Harvard, T.H. Chan School of Public Health, 10Cardiovascular Division, Washington University School of Medicine, Washington University, 11Centro de Estudios en Salud, Universidad del Valle de Guatemala, 12Division of Pulmonary and Critical Care, School of Medicine, Johns Hopkins University

We detail the consistent, high-quality procedures used throughout air and biological sampling processes at Indian field sites during a large randomized controlled trial. Insights gathered from the oversight of applications of innovative technologies, adapted for exposure assessment in rural regions, enable better field data collection practices with more reliable outcomes.

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Biology

Compost Microcosms as Microbially Diverse, Natural-like Environments for Microbiome Research in Caenorhabditis elegans
Kenneth Trang 1, Rahul Bodkhe 1, Michael Shapira 1
1Department of Integrative Biology, University of California, Berkeley

Compost microcosms bring the microbial diversity found in nature into the laboratory to facilitate microbiome research in Caenorhabditis elegans. Provided here are protocols for setting up microcosm experiments, with the experiments demonstrating the ability to modulate environmental microbial diversity to explore the relationships between environmental microbial diversity and worm gut microbiome composition.

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Bioengineering

Mechano-Node-Pore Sensing: A Rapid, Label-Free Platform for Multi-Parameter Single-Cell Viscoelastic Measurements
Andre Lai *1, Rachel Rex *2, Kristen L. Cotner 1, Alan Dong 3, Michael Lustig 1,3, Lydia L. Sohn 1,2
1Graduate Program in Bioengineering, University of California, Berkeley and University of California, San Francisco, 2Department of Mechanical Engineering, University of California, Berkeley, 3Department of Electrical Engineering and Computer Sciences, University of California, Berkeley

Presented here is a method to mechanically phenotype single cells using an electronics-based microfluidic platform called mechano-node-pore sensing (mechano-NPS). This platform maintains moderate throughput of 1-10 cells/s while measuring both the elastic and viscous biophysical properties of cells.

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Biology

Isolation of Retinal Pigment Epithelial Cells from Guinea Pig Eyes
So Goto 1,2,3, Michael Frost 4, Christine Wildsoet 1
1Herbert Wertheim School Optometry and Vision Science, University of California, Berkeley, 2Department of Ophthalmology, Osaka University Graduate School of Medicine, 3Department of Ophthalmology, Tokyo Medical Center, National Hospital Organization, 4Department of Optometry and Vision Science, School of Optometry, University of Alabama at Birmingham

We describe a simple and efficient method for isolating cells of the retinal pigment epithelium (RPE) cells from the eyes of young pigmented guinea pigs. This procedure allows for follow-up molecular biology studies on the isolated RPE, including gene expression analyses.

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Neuroscience

Bringing the Clinic Home: An At-Home Multi-Modal Data Collection Ecosystem to Support Adaptive Deep Brain Stimulation
Gabrielle Strandquist 1, Tomasz Frączek 2, Tanner Dixon 3, Shravanan Ravi 3, Raphael Bechtold 4, Daryl Lawrence 5, Alicia Zeng 6, Jack Gallant 7, Simon Little 3, Jeffrey Herron 8
1Computer Science and Engineering, University of Washington, 2Neuroscience, University of Washington, 3Neurology, University of California, San Francisco, 4Bioengineering, University of Washington, 5Bioengineering, University of California, Berkeley, 6Biophysics, University of California, Berkeley, 7Psychology, University of California, Berkeley, 8Neurological Surgery, University of Washington

The protocol shows a prototype of the at-home multi-modal data collection platform that supports research optimizing adaptive deep brain stimulation (aDBS) for people with neurological movement disorders. We also present key findings from deploying the platform for over a year to the home of an individual with Parkinson's disease.

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Biology

Transgene Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors
Brian Benyamini 1, Meagan N. Esbin 1,2, Oscar Whitney 1, Nike Walther 1, Anna C. Maurer 1
1Department of Molecular and Cell Biology, University of California, Berkeley, 2Biophysics Graduate Group, University of California, Berkeley

Recombinant adeno-associated virus (rAAV) is widely used for clinical and preclinical gene delivery. An underappreciated use for rAAVs is the robust transduction of cultured cells without the need for purification. For researchers new to rAAV, we provide a protocol for transgene cassette cloning, crude vector production, and cell culture transduction.

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Biochemistry

Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation
Trinity Cookis 1, Paul Sauer 2,3, Simon Poepsel 4,5, Bong-Gyoon Han 6, Dominik A. Herbst 1,2,6, Robert Glaeser 6, Eva Nogales 1,2,3,6
1Department of Molecular and Cell Biology, University of California, Berkeley, 2California Institute for Quantitative Biology (QB3), University of California, Berkeley, 3Howard Hughes Medical Institute, University of California, Berkeley, 4Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital, University of Cologne, 5Cologne Excellence Cluster for Cellular Stress Responses in Ageing-Associated Diseases (CECAD), University of Cologne, 6Molecular Biophysics and Integrative Bio-Imaging Division, Lawrence Berkeley National Laboratory

A step-by-step protocol for fabricating streptavidin affinity grids is provided for use in structural studies of challenging macromolecular samples by cryo-electron microscopy.

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Cancer Research

Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model
Kerem Nernekli *1, Dilyana B. Mangarova *1, Yifeng Shi 2, Zahra Shokri Varniab 1, Edwin Chang 1, Oguz Ziya Tikenogullari 3, Laura Pisani 1, Grigory Tikhomirov 2, Gordon Wang 4, Heike E. Daldrup-Link 1
1Molecular Imaging Program at Stanford (MIPS), Department of Radiology, Stanford University School of Medicine, 2Department of Electrical Engineering and Computer Sciences, University of California, Berkeley, 3Department of Mechanical Engineering, Stanford University, 4Department of Psychiatry and Behavioral Sciences, Stanford University, Wu Tsai Neuroscience Institute, Stanford University

We present a novel approach for two-photon microscopy of the tumor delivery of fluorescent-labeled iron oxide nanoparticles to glioblastoma in a mouse model.

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