Physical Sciences Tutorials

X40 - Advanced Cryo-FIB Specimen Preparation and Handling of Environmentally Sensitive Materials for APT and TEM Analysis

PRESENTER:
Daniel Perea, Pacific Northwest National Laboratory

Dr. Perea is a senior staff scientist at the Environmental Molecular Sciences Laboratory at the Pacific Northwest National Laboratory. He received his Ph.D. in materials science & engineering from Northwestern University in 2009. Dr. Perea has over 14 years of experience using atom probe tomography, with much of his expertise in semiconductor and structural materials. His current interests at PNNL include developing unique cryogenic-based hardware and protocols to pioneer the use of atom probe tomography to probe the composition and structure of hydrated materials.

• Applications of APT to hydrated interfaces such as those in biological and environmental materials
• Development of a site-specific liftout and attachment scheme for cryogenically cooled specimens
• Challenges of handling and transfer of post cryo-prepared specimens between tools.
• Specimen shuttle suitcase devices for transfer of environmentally sensitive materials between the FIB and other tools enabling APT and TEM analysis

X41 - Entrepreneurship in the Microscopy Community

PRESENTER:
Benjamin Bammes, Direct Electron

Benjamin has a broad scientific background, including physics, computer science, biology, and chemistry, working with small businesses related to science since 2003 in addition to academic research. He has a Ph.D. in Molecular Biophysics from Baylor College of Medicine, where he worked under National Academy of Sciences member Dr. Wah Chiu to optimize cryo-EM data collection and image processing. In 2011, Benjamin joined Direct Electron and now leads our collaborative approach to delivering high-tech solutions to the scientific community.

Several entrepreneurs from the microscopy community participate in an open Q&A forum on topics related to starting and running a microscopy-related business. Topics may include:

• How to identify and develop potential new products
• Steps for successful commercialization
• Methods for funding the early stages of a business
• Marketing and gaining initial customers
• Protecting and leveraging intellectual property
• The role of academic collaborations

X42 - TriBeam Tomography for 3D Data Acquisition

PRESENTER:
McLean Echlin, University of California-Santa Barbara

McLean Echlin is a research scientist at UC-Santa Barbara where his interests are in problems related to 3D materials science, strain mapping, and other advanced characterization techniques that are applicable at the mesoscale. Over the past decade he has developed multiple iterations of the femtosecond laser enabled TriBeam microscope for mm-scaled 3D measurements in a wide range of materials.

In this tutorial, the femtosecond laser enabled TriBeam microscope for use in tomography and bulk sample excavation is discussed, which will include the following topics:

• Femtosecond laser material interactions during serial sectioning
• Multimodal data acquisition and experimental setup
• Automation and dynamically varying experiment parameters during tomography
• Reconstruction and image processing of 3D data

Biological Sciences Tutorials

X43 - Liquid Cell TEM Imaging Techniques and Optimization for Biological Samples

PRESENTER:
Madeline Dukes, Protochips, Inc.

Madeline Dukes is an applications scientist with Protochips, Inc. She has more than 10 years of experience in the field of liquid-cell transmission electron microscopy (LC-TEM) and has worked with multiple labs around the world to improve in-situ LC-TEM technology and develop new sample preparation techniques for a broad range of applications.

• Liquid Cell Transmission Electron Microscopy (LC-TEM) for biological systems — challenges & applications
• Strategies to minimize beam and radiolysis damage when imaging biological samples using LC-TEM
• Selecting the best parameters for imaging biological samples in liquid, including E-chip selection, flow conditions, etc.
• Sample preparation workflows for optimizing LC-TEM experiments, such as affinity-coating, staining, cell culture etc.

X44 - Optimization of Cryo-EM Data Collection using Advanced Direct Detectors

PRESENTER:
Michael Spilman, Direct Electron

Michael is broadly interested in applying novel imaging methods using direct electron detectors to bridge high resolution biological structures with real-time dynamics. At Direct Electron, he supports and collaborates with customers on numerous projects including single particle cryoEM, microED, and liquid cell in situ.

• Optimizing CryoEM data collection for direct electron detectors
• When to use counting and linear modes for certain experiments
• Discussion of single particle, tomography, microED, and additional methods using direct electron detectors

X45 - CryoEM Sample Preparation: Problems and Potential Solutions

PRESENTER:
Alex Noble, New York Structural Biology Center

Alex is broadly interested in developing and applying methods for solving biomedical problems. He is the primary tomographer at SEMC. He works on several projects including cryo-FIB/SEM, tomography of cells, reconstituted systems, purified protein samples, tomography software development, and deep learning applications in cryoEM.

• What potential air-water interface issues exist in cryo-EM/ET?
• Why is preferred orientation an issue?
• How may air-water interface issues be reduced or avoided?
• How may preferred orientation issues be ameliorated?
• What are the basics for preparing cell samples for cryo-FIB/SEM?