Lawrence Berkeley National Laboratory

33 ARTICLES PUBLISHED IN JoVE

Biology

Label-free in situ Imaging of Lignification in Plant Cell Walls

Martin Schmidt¹, Pradeep Perera¹, Adam M. Schwartzberg², Paul D. Adams³, P. James Schuck²

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.

Bioengineering

Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets

Helen Tran¹, Sarah L. Gael¹, Michael D. Connolly¹, Ronald N. Zuckermann¹

¹Molecular Foundry, Lawrence Berkeley National Laboratory

A simple and general manual peptoid synthesis method involving basic equipment and commercially available reagents is outlined, enabling peptoids to be easily synthesized in most laboratories. The synthesis, purification and characterization of an amphiphilic peptoid 36mer is described, as well as its self-assembly into highly-ordered nanosheets.

Biology

Fabrication and Use of MicroEnvironment microArrays (MEArrays)

Chun-Han Lin^1,2, Jonathan K. Lee¹, Mark A. LaBarge¹

¹Life Science Division, Lawrence Berkeley National Laboratory, ²Department of Comparative Biochemistry, University of California, Berkeley

A combinatorial functional screening method for gaining insights into the impacts of the molecular composition of microenvironments on cellular functions is described. The method takes advantage of existing microarray-based technologies to generate arrays of defined combinatorial microenvironments that support cell adhesion and functional analysis.

Medicine

Processing of Human Reduction Mammoplasty and Mastectomy Tissues for Cell Culture

Mark A. LaBarge¹, James C. Garbe¹, Martha R. Stampfer¹

¹Life Science Division, Lawrence Berkeley National Laboratory

A method to process human mammary surgical discard material is described. Processed tissue, in the form of organoids, can be stored frozen indefinitely or placed in culture for long-term growth. This method enables experimental examination of normal human epithelial cell biology, and the effects of exogenous perturbations.

Engineering

Revealing Dynamic Processes of Materials in Liquids Using Liquid Cell Transmission Electron Microscopy

Kai-Yang Niu¹, Hong-Gang Liao¹, Haimei Zheng¹

¹Materials Sciences Division, Lawrence Berkeley National Laboratory

We have developed a self-contained liquid cell, which allows imaging through liquids using a transmission electron microscope. Dynamic processes of nanoparticles in liquids can be revealed in real time with sub-nanometer resolution.

JoVE Journal

Using High Resolution Computed Tomography to Visualize the Three Dimensional Structure and Function of Plant Vasculature

Andrew J. McElrone^1,2, Brendan Choat³, Dilworth Y. Parkinson⁴, Alastair A. MacDowell⁴, Craig R. Brodersen⁵

¹U.S. Department of Agriculture, ²Department of Viticulture and Enology, University of California - Davis, ³Hawkesbury Institute for the Environment, University of Western Sydney, ⁴Advanced Light Source, Lawrence Berkeley National Lab, ⁵Citrus Research & Education Center, University of Florida

High resolution x-ray computed tomography (HRCT) is a non-destructive diagnostic imaging technique that can be used to study the structure and function of plant vasculature in 3D. We demonstrate how HRCT facilitates exploration of xylem networks across a wide range of plant tissues and species.

Engineering

Molecular Beam Mass Spectrometry With Tunable Vacuum Ultraviolet (VUV) Synchrotron Radiation

Amir Golan¹, Musahid Ahmed¹

¹Chemical Sciences Division, Lawrence Berkeley National Laboratory

A molecular beam coupled to tunable vacuum ultraviolet photoionization mass spectrometer at a synchrotron provides a convenient tool to explore the electronic structure of isolated gas phase molecules and clusters. Proton transfer mechanisms in DNA base dimers were elucidated with this technique.

Engineering

Characterization of Electrode Materials for Lithium Ion and Sodium Ion Batteries Using Synchrotron Radiation Techniques

Marca M. Doeff¹, Guoying Chen¹, Jordi Cabana^1,2, Thomas J. Richardson¹, Apurva Mehta³, Mona Shirpour¹, Hugues Duncan¹, Chunjoong Kim¹, Kinson C. Kam⁴, Thomas Conry⁵

¹Environmental Energy Technologies Division, Lawrence Berkeley National Laboratory, ²Department of Chemistry, University of Illinois at Chicago, ³Stanford Synchrotron Radiation Lightsource, ⁴Haldor Topsøe A/S, ⁵PolyPlus Battery Company

We describe the use of synchrotron X-ray absorption spectroscopy (XAS) and X-ray diffraction (XRD) techniques to probe details of intercalation/deintercalation processes in electrode materials for Li-ion and Na-ion batteries. Both in situ and ex situ experiments are used to understand structural behavior relevant to the operation of devices

Chemistry

Seeded Synthesis of CdSe/CdS Rod and Tetrapod Nanocrystals

Karthish Manthiram^*1,4, Brandon J. Beberwyck^*2,4, Dmitri V. Talapin^5,6, A. Paul Alivisatos^2,3,4

¹Department of Chemical Engineering, UC Berkeley, ²Department of Materials Science and Engineering, UC Berkeley, ³Department of Chemistry, UC Berkeley, ⁴Materials Sciences Division, Lawrence Berkeley National Laboratory, ⁵Department of Chemistry, University of Chicago, ⁶Center for Nanoscale Materials, Argonne National Laboratory

A protocol for the seeded synthesis of rod-shaped and tetrapod-shaped multicomponent nanostructures consisting of CdS and CdSe is presented.

Engineering

Flame Experiments at the Advanced Light Source: New Insights into Soot Formation Processes

Nils Hansen¹, Scott A. Skeen¹, Hope A. Michelsen¹, Kevin R. Wilson², Katharina Kohse-Höinghaus³

¹Combustion Research Facility, Sandia National Laboratories, ²Chemical Sciences Division, Advanced Light Source, Lawrence Berkeley National Laboratory, ³Physikalische Chemie I, Universität Bielefeld

Gas sampling from laboratory-scale flames with online analysis of all species by mass spectrometry is a powerful method to investigate the complex mixture of chemical compounds occurring during combustion processes. Coupled with tunable soft ionization via synchrotron-generated vacuum-ultraviolet radiation, this technique provides isomer-resolved information and potentially fragment-free mass spectra.

Biology

Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements

Prajakta Pradhan Mitra^1,2, Dominique Loqué^1,2

¹Feedstocks Division, Joint Bioenergy Institute, ²Physical Biosciences Division, Lawrence Berkeley National Laboratory

Plant cell wall composition varies between tissue types and can include lignin, cellulose, hemicelluloses, and pectin. Various staining techniques have been developed to visualize differences at the cell-type level. This paper is a compilation of commonly used cell wall staining techniques.

Bioengineering

Lipid Bilayer Vesicle Generation Using Microfluidic Jetting

Christopher W. Coyne¹, Karan Patel¹, Johanna Heureaux¹, Jeanne Stachowiak³, 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.

Bioengineering

From Voxels to Knowledge: A Practical Guide to the Segmentation of Complex Electron Microscopy 3D-Data

Wen-Ting Tsai¹, Ahmed Hassan¹, Purbasha Sarkar², Joaquin Correa^1,3, Zoltan Metlagel¹, Danielle M. Jorgens¹, Manfred Auer^1,2

¹Life Sciences Division, Lawrence Berkeley National Laboratory, ²Joint Bioenergy Institute, Physical Biosciences Division, Lawrence Berkeley National Laboratory, ³National Energy Research Scientific Computing Center, Lawrence Berkeley National Laboratory

The bottleneck for cellular 3D electron microscopy is feature extraction (segmentation) in highly complex 3D density maps. We have developed a set of criteria, which provides guidance regarding which segmentation approach (manual, semi-automated, or automated) is best suited for different data types, thus providing a starting point for effective segmentation.

Biology

DNA-affinity-purified Chip (DAP-chip) Method to Determine Gene Targets for Bacterial Two component Regulatory Systems

Lara Rajeev¹, Eric G. Luning¹, Aindrila Mukhopadhyay¹

¹Physical Biosciences Division, Lawrence Berkeley National Laboratory

This video article describes an in vitro microarray based method to determine the gene targets and binding sites for two component system response regulators.

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.

Engineering

Fabrication of Gate-tunable Graphene Devices for Scanning Tunneling Microscopy Studies with Coulomb Impurities

Han Sae Jung^1,2, Hsin-Zon Tsai¹, Dillon Wong¹, Chad Germany¹, Salman Kahn¹, Youngkyou Kim^1,3, Andrew S. Aikawa¹, Dhruv K. Desai¹, Griffin F. Rodgers¹, Aaron J. Bradley¹, Jairo Velasco Jr.¹, Kenji Watanabe⁴, Takashi Taniguchi⁴, Feng Wang^1,5,6, Alex Zettl^1,5,6, Michael F. Crommie^1,5,6

¹Department of Physics, University of California at Berkeley, ²Department of Chemistry, University of California at Berkeley, ³Department of Chemical and Biomolecular Engineering, University of California at Berkeley, ⁴National Institute for Materials Science (Japan), ⁵Materials Sciences Division, Lawrence Berkeley National Laboratory, ⁶Kavli Energy NanoSciences Institute, University of California at Berkeley and Lawrence Berkeley National Laboratory

This paper details the fabrication process of a gate-tunable graphene device, decorated with Coulomb impurities for scanning tunneling microscopy studies. Mapping the spatially dependent electronic structure of graphene in the presence of charged impurities unveils the unique behavior of its relativistic charge carriers in response to a local Coulomb potential.

Engineering

Failure Analysis of Batteries Using Synchrotron-based Hard X-ray Microtomography

Katherine J. Harry^1,2, Dilworth Y. Parkinson³, Nitash P. Balsara^2,4,5

¹Department of Materials Science and Engineering, University of California Berkeley, ²Materials Science Division, Lawrence Berkeley National Laboratory, ³Advanced Light Source Division, Lawrence Berkeley National Laboratory, ⁴Department of Chemical and Biomolecular Engineering, University of California Berkeley, ⁵Environmental Energy Technology Division, Lawrence Berkeley National Laboratory

Synchrotron-based hard X-ray microtomography is used to image the electrochemical growth of dendrites from a lithium metal electrode through a solid polymer electrolyte membrane.

Neuroscience

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

Feng Liu¹, Nannan Liu¹

¹Department of Entomology and Plant Pathology, Auburn University

Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

Engineering

Using Synchrotron Radiation Microtomography to Investigate Multi-scale Three-dimensional Microelectronic Packages

Holly D. Carlton¹, John W. Elmer¹, Yan Li², Mario Pacheco², Deepak Goyal², Dilworth Y. Parkinson³, Alastair A. MacDowell³

¹Materials Engineering Division, Lawrence Livermore National Laboratory, ²Assembly Test and Technology Development Failure Analysis Labs, Intel Corporation, ³Advanced Light Source, Lawrence Berkeley National Laboratory

For this study synchrotron radiation micro-tomography, a non-destructive three-dimensional imaging technique, is employed to investigate an entire microelectronic package with a cross-sectional area of 16 x 16 mm. Due to the synchrotron's high flux and brightness the sample was imaged in just 3 min with an 8.7 µm spatial resolution.

JoVE Core

Printing Fabrication of Bulk Heterojunction Solar Cells and In Situ Morphology Characterization

Feng Liu¹, Sunzida Ferdous², Xianjian Wan¹, Chenhui Zhu³, Eric Schaible³, Alexander Hexemer³, Cheng Wang³, Thomas P. Russell^1,2

¹Materials Sciences Division, Lawrence Berkeley National Laboratory, ²Department of Polymer Science and Engineering, University of Massachusetts, Amherst, ³Advanced Light Source, Lawrence Berkeley National Laboratory

Here, we present a protocol to fabricate organic thin film solar cells using a mini-slot die coater and related in-line structure characterizations using synchrotron scattering techniques.

Biology

Experimental Approach for Determining Semiconductor/liquid Junction Energetics by Operando Ambient-Pressure X-ray Photoelectron Spectroscopy

Michael Frankston Lichterman^*1, Matthias H Richter^*2, Shu Hu^*1, Ethan J Crumlin^*3, Stephanus Axnanda³, Marco Favaro³, Walter Drisdell³, Zahid Hussein³, Bruce S Brunschwig⁴, Nathan S Lewis¹, Zhi Liu³, Hans-Joachim Lewerenz²

¹Division of Chemistry and Chemical Engineering, California Institute of Technology, ²Joint Center for Artificial Photosynthesis, California Institute of Technology, ³Advanced Light Source, Lawrence Berkeley National Laboratory, ⁴Beckman Institute, California Institute of Technology

We present a method for the determination of the energy relations of semiconductor/liquid junctions, which are the basis for the successful operation of such renewable solar energy converting systems.

Chemistry

Synthesis of a Water-soluble Metal–Organic Complex Array

Purnandhu Bose¹, Pradip K. Sukul¹, Omar M. Yaghi^2,3,4, Kentaro Tashiro¹

¹International Center for Materials Nanoarchitectonics, National Institute for Materials Science, ²Department of Chemistry, University of California–Berkeley, ³Materials Sciences Division, Lawrence Berkeley National Laboratory, ⁴Kavli Energy NanoSciences Institute at Berkeley, University of California–Berkeley

A potential general method for the synthesis of water-soluble multimetallic peptidic arrays containing a predetermined sequence of metal centers is presented.

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. Sharp¹, Francesca M. Toma¹

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 CH₃NH₃PbI_3-xBr_x thin films for the application in high efficiency solar cells and other optoelectronic devices.

Biochemistry

Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE

Venkataramana R. Pidatala^*1,2, Amir Mahboubi^*1,2, Jenny C. Mortimer^1,2

¹Joint BioEnergy Institute, ²Environmental and Systems Biology, BioSciences Division, Lawrence Berkeley National Laboratory

A protocol for the structural analysis of polysaccharides by gel electrophoresis (PACE), using mannan as an example, is described.

Developmental Biology

Application of Aorta-gonad-mesonephros Explant Culture System in Developmental Hematopoiesis

Junhua Lv^1,2, Feng Liu^1,2

¹State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, ²University of Chinese Academy of Sciences

This protocol describes using cultured Aorta-Gonad-Mesonephros for expression analyses, colony-forming units in the culture and spleen, and long-term reconstitution to determine the effect of regulatory factors and signaling pathways on hematopoietic stem cell development. This has been demonstrated as an effective system for studying hematopoietic stem cell biology and function.

JoVE Core

Electric-field Control of Electronic States in WS₂ Nanodevices by Electrolyte Gating

Feng Qin¹, Toshiya Ideue¹, Wu Shi², Yijin Zhang^3,4, Ryuji Suzuki¹, Masaro Yoshida⁵, Yu Saito¹, Yoshihiro Iwasa^1,5

¹Quantum-Phase Electronics Center (QPEC) and Department of Applied Physics, The University of Tokyo, ²Materials Sciences Division, Lawrence Berkeley National Laboratory, ³Institute of Scientific and Industrial Research, Osaka University, ⁴Max Planck Institute for Solid State Research, ⁵RIKEN Center for Emergent Matter Science (CEMS)

Here, we present a protocol to control the carrier number in solids by using the electrolyte.

Environment

Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

Jian Gao^1,2, Joelle Sasse^1,2, Kyle M. Lewald^1,2, Kateryna Zhalnina^1,2, Lloyd T. Cornmesser^1,2, Todd A. Duncombe³, Yasuo Yoshikuni², John P. Vogel², Mary K. Firestone⁴, Trent R. Northen^1,2

¹Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, ²Joint Genome Institute, Department of Energy, ³Joint BioEnergy Institute, ⁴Department of Environmental Science Policy and Management, University of California

This article describes detailed protocols for ecosystem fabrication of devices (EcoFABs) that enable the studies of plants and plant-microbe interactions in highly controlled laboratory conditions.

Chemistry

Elemental-sensitive Detection of the Chemistry in Batteries through Soft X-ray Absorption Spectroscopy and Resonant Inelastic X-ray Scattering

Jinpeng Wu^1,2, Shawn Sallis^2,3, Ruimin Qiao², Qinghao Li^2,4, Zengqing Zhuo^2,5, Kehua Dai^2,6, Zixuan Guo^2,7, Wanli Yang²

¹Geballe Laboratory for Advanced Materials, Stanford University, ²Advanced Light Source, Lawrence Berkeley National Laboratory, ³Department of Materials Science and Engineering, Binghamton University, ⁴School of Physics, National Key Laboratory of Crystal Materials, Shandong University, ⁵School of Advanced Materials, Peking University Shenzhen Graduate School, ⁶School of Metallurgy, Northeastern University, ⁷Department of Chemical Engineering, University of California-Santa Barbara

Here, we present a protocol for typical experiments of soft X-ray absorption spectroscopy (sXAS) and resonant inelastic X-ray scattering (RIXS) with applications in battery material studies.

Chemistry

Using Graphene Liquid Cell Transmission Electron Microscopy to Study in Situ Nanocrystal Etching

Matthew R. Hauwiller¹, Justin C. Ondry¹, A. Paul Alivisatos^1,2,3,4

¹Department of Chemistry, University of California-Berkeley, ²Department of Material Science and Engineering, University of California-Berkeley, ³Kavli Energy NanoScience Institute, ⁴Materials Sciences Division, Lawrence Berkeley National Laboratory

Graphene liquid cell electron microscopy can be used to observe nanocrystal dynamics in a liquid environment with greater spatial resolution than other liquid cell electron microscopy techniques. Etching premade nanocrystals and following their shape using graphene liquid cell Transmission Electron Microscopy can yield important mechanistic information about nanoparticle transformations.

JoVE Journal

Synchrotron X-ray Microdiffraction and Fluorescence Imaging of Mineral and Rock Samples

Camelia V. Stan¹, Nobumichi Tamura¹

¹Advanced Light Source, Lawrence Berkeley National Laboratory

We describe a beamline setup meant to carry out rapid two-dimensional x-ray fluorescence and x-ray microdiffraction mapping of single crystal or powder samples using either Laue (polychromatic radiation) or powder (monochromatic radiation) diffraction. The resulting maps give information about strain, orientation, phase distribution, and plastic deformation.

Medicine

Real-Time Dynamic Navigation System for the Precise Quad-Zygomatic Implant Placement in a Patient with a Severely Atrophic Maxilla

Yihan Shen^*1, Qinggang Dai^*1, Baoxin Tao^*1, Wei Huang², Feng Wang², Kengliang Lan¹, Yuanyuan Sun¹, Xiaowan Ling¹, Lijun Yan¹, Yueping Wang¹, Yiqun Wu¹

¹The 2nd Dental Center, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, ²Department of Dental Implantology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology

Here, we present a protocol to achieve precise quad-zygomatic implant placement in patients with severely atrophic maxilla using a real-time dynamic navigation system.

Environment

Bioprospecting of Extremophilic Microorganisms to Address Environmental Pollution

Giovanni Gallo^1,2, Martina Aulitto^1,3, Patrizia Contursi^1,4, Danila Limauro^1,4, Simonetta Bartolucci¹, Gabriella Fiorentino^1,4

¹Department of Biology, University of Naples Federico II, ²Institute of Polymers, Composites and Biomaterials (IPCB), Consiglio Nazionale delle Ricerche CNR, ³Division of Biological Systems and Engineering, Lawrence Berkeley National Laboratory, ⁴BAT Center-Interuniversity Center for Studies on Bioinspired Agro-Environmental Technology, University of Napoli Federico II

The isolation of heavy metal-resistant microbes from geothermal springs is a hot topic for the development of bioremediation and environmental monitoring biosystems. This study provides a methodological approach for isolating and identifying heavy metal tolerant bacteria from hot springs.

Biochemistry

Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation

Trinity Cookis¹, Paul Sauer^2,3, Simon Poepsel^4,5, Bong-Gyoon Han⁶, Dominik A. Herbst^1,2,6, Robert Glaeser⁶, 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.