You are invited to the New England Society for Microscopy's Virtual Fall Symposium! The meeting will take place on November 20th via Zoom, consisting of four technical talks, four lightning talks, our Annual Business Meeting, and an afternoon workshop on using ImageJ given by Lai Ding of Harvard Medical School. The technical lectures will be delivered by Madeline Oudin (Tufts), Guillermina Ramirez-San Juan (Brandeis), Arthur McClelland (Harvard), and Eddie Pang (Schuh Lab, MIT). The lightning talks will be delivered by Linda Wiratan (Harvard University), Sandra A. McDonald (Staff Pathologist, John J. Godleski MD PLLC), Stephan Goodman (Microscopy Innovations, LLC) and Jules A. Gardener (Center for Nanoscale Systems, Harvard).
Please register (for FREE!) to receive the Zoom link. Log on at 8:45 am, talks will begin at 9 am.
Meeting Schedule, November 20th
8:45 AM – Zoom Meeting will open
9:00 AM – Guillermina Ramirez-San Juan (Brandeis)
How do organisms pattern fluid flows?
9:40 AM – Arthur McClelland (Harvard)
Probing historic photographs with FTIR microscopy
10:20 AM – Business Meeting
10:50 AM – Eddie Pang (Schuh Lab, MIT)
Overcoming difficulties with pseudo symmetry in electron backscatter diffraction: a case study in zirconia shape-memory ceramics
11:30 AM - Madeline Oudin (Tufts)
Using in vitro and in vivo microscopy approaches to dissect the roles of the extracellular matrix in cancer metastasis and chemoresistance
12:10 PM - Break for lunch
1:15 PM - Lightning talks
1:15 PM Linda Wiratan (Harvard)
Live-cell and TEM Imaging of Gently Preserved Tissues using a Novel Biopsy-Freezing Technique
1:25 PM Sandra A. McDonald
Scanning Electron Microscopic Detection of Talc and Fibrous Particles in the Female Genital Tract
1:35 PM Stephan Goodman (Microscopy Innovations, LLC)
Automated and Very Fast Preparation of Biological Specimens for Transmission and 3D Volume Electron Microscopy
1:45 PM Jules Gardener (Center for Nanoscale Systems, Harvard)
Using a high-speed direct detection camera for ultrafast 4D-STEM data collection
2:00 PM - ImageJ Workshop- Given by Lai Ding (Harvard Medical School)
All participants should have FIJI ImageJ installed and ready to use on their computers and have the folder from Lai Ding (which will be emailed to registrants on Thursday) ready to open on their desktop.
ABSTRACTS AND BIOS
Guillermina Ramirez-San Juan (Brandeis)
Title: How do organisms pattern fluid flows?
Abstract:The generation of flow is central to the development and physiology of organisms across the tree of life. Flows are responsible for functions that range from feeding and swimming in single cell protists to mucus clearance in humans. These flows are generated by the activity of thousands of micrometer scale cilia anchored to the cell surface that beat at frequencies of 15-50Hz. The lack of experimental studies where cilia, fluid and their interactions are imaged across all the relevant spatio-temporal scales has hindered an integrated study of flow generation. In this talk I will show how live imaging of a variety of model organisms can help us identify the key physical parameters of individual cilia (e.g. cilia beat frequency) that control the emergent properties of a cilia array (e.g. beat synchronization) and flow (e.g. structure). In particular, I will discuss how imaging of the spatial distribution and activity of cilia in the mouse airway from the sub-cellular (nm) to the organ scales (cm) has advanced our understanding of mucus clearance. Altogether this approach will allow us to link the structure of the flow generated by a ciliary array with its underlying spatial organization. This framework is specially valuable to understand the origin of defects in fluid transport, which are the underlying cause of human pathologies.
Arthur McClelland (Harvard)
Title: Probing historic photographs with FTIR microscopy
Abstract:
Fourier Transform Infrared (FTIR) Microscopy allows for nondestructive chemical identification of samples. Here I will present a project in collaboration with the Weissman Preservation Center at Harvard Library on identifying the coatings on historic salt print photographs. I will review FTIR microscopy for those who are not familiar with the technique. I will also quickly review the early invention of photography which will hopefully be of interest to microscopists. Then I will share the results of the analysis of the original Harvard yearbook from 1853. Time allowing, I will conclude with some interesting future directions of the project has started to take me.
Biography:
Dr. Arthur McClelland received an Associate’s degree from Simon’s Rock College of Bard, a Bachelor’s degree in Engineering Physics from the University of Pittsburgh, a Master’s degree in Electrical Engineering from the University of Michigan, and a Doctorate degree in Applied Physics also from the University of Michigan. He did his postdoctoral research at Northeastern University, before joining the technical staff at the Center for Nanoscale Systems at Harvard University in 2011 where he runs the optical spectroscopy and microscopy lab. Arthur was a physical sciences director for NESM from 2011 - 2014. He is a co-author of Optical Measurements for Scientists and Engineers: A Practical Guide from Cambridge University Press which was published in 2018. He has also periodically co-taught microscopy courses at the Harvard Extension School.
Madeleine Oudin PhD, Tufts University, BME
Title: Using in vitro and in vivo microscopy approaches to dissect the roles of the extracellular matrix in cancer metastasis and chemoresistance
Abstract:
The extracellular matrix (ECM) is a major component of the tumor microenvironment, where it can support cellular growth, promote local invasion from the primary tumor, and contribute to metastatic outgrowth in distant sites. Our research focuses on understanding how individual ECM proteins upregulated in metastatic breast tumors drive local invasion and support metastasis. We use a range of in vitro models that replicate the ECM organization found in vivo to dissect mechanisms of cell migration in vitro and the signaling pathways activated by the ECM. Further, we have used intravital imaging in breast tumors to dissect the local invasion of tumor cells in vivo in response to individual cues or after treatment responses. Overall, our studies shed light on novel ways in which the ECM drives invasion and chemoresistance.
Biography:
Dr. Oudin completed a BSc in Biochemistry at McGill University and a PhD in Neuroscience from King’s College London, UK, where she studied the regulation of neuronal cell migration. As a post-doctoral fellow in Prof. Frank Gertler’s lab at the Koch Institute for Integrative Cancer Research at MIT, her research was focused on understanding how metastasis, the dissemination of tumor cells throughout the body, occurs and how aggressive cells become resistant to current treatments. She received a Breast Cancer Research Department of Defence Post-doctoral Fellowship and a K99/R00 Pathway to Independence from the National Cancer Institute. She started as an Assistant Professor at Tufts University in the department of Biomedical Engineering in 2018, where her lab is focused on understanding the mechanisms by which the tumor microenvironment contributes to cancer metastasis and resistance to drugs. She has also received multiple awards, such as the Women in Cancer Research Award, American Association for Cancer Research Scholar-in-Training Award, the MIT Infinite Kilometer Award and CMBE Rising Star Award.
Edward L. Pang and Christopher A. Schuh (MIT)
Title: Overcoming difficulties with pseudosymmetry in electron backscatter diffraction: a case study in zirconia shape-memory ceramics
Abstract:
Electron backscatter diffraction (EBSD) is a popular characterization technique used to measure the crystallographic orientation of crystalline materials. However, current EBSD systems struggle to correctly index pseudosymmetric materials such as the tetragonal phase of zirconia shape-memory ceramics. Here, we present a new way to analyze EBSD data to solve this problem, a modified dictionary indexing approach. We demonstrate that the enhanced sensitivity of this method can successfully and efficiently resolve pseudosymmetry in both simulated and experimental patterns of tetragonal zirconia. We then highlight the importance of this advance to our group’s recent work in understanding the role of phase transformation crystallography on the properties of zirconia shape-memory ceramics.
Biography:
Edward Pang is a 5th year Ph.D. student in Materials Science & Engineering at MIT and is an NSF Graduate Research Fellow, Churchill Scholar, and Goldwater Scholar. His research focuses on the role of crystallography on the functional properties of shape-memory alloys and ceramics as well as the development of new methods to enhance the capabilities of the electron backscatter diffraction technique. He has authored 9 publications in peer-reviewed journals.
ImageJ Workshop with Lai Ding:
The workshop will cover basic Image analysis operation including: open image with various format, save image; image thresholding/segmentation, ROI and measurement, cell identification manual+auto.
All participants should have FIJI ImageJ installed and ready to use on their computers and have the folder from Lai Ding (which will be emailed to registrants on Thursday) ready to open on their desktop.
Biography:
Dr. Ding, the senior scientist of NeuroTehcnoology Studio (NTS) has solid knowledge and expertise in optical imaging techniques and digital data processing. His background in Physics and mathematics allows exceptional command of the theories and implementation of modern optical imaging. His 16 years of experiences as the manager of optical imaging core facilities collaborating with numerous researchers has led to in depth understanding of the practical application of optical imaging in biology.
Lightning Talk Abstracts:
Linda Wiratan (Harvard University)
Live-cell and TEM Imaging of Gently Preserved Tissues using a Novel Biopsy-Freezing Technique
Techniques such as CLEM, which integrate the dynamics of live-cell microscopy with the high resolution of TEM, are powerful but limited by sample quality. These samples usually experience chemical/physical stress during collection, so their ultrastructure can deviate from the native structure, especially for tissues from a live organism. I am engineering a novel biopsy-freezing technique to cleanly excise out tissue cores with biopsy and rapidly freeze them under cryogen. The resulting tissue will have gently-preserved three-dimensional ultrastructure. I am currently characterizing ice formation within the biopsy needle to optimize vitrification quality.
Sandra A. McDonald
Scanning Electron Microscopic Detection of Talc and Fibrous Particles in the Female Genital Tract
Analyses on human tissue are important to the discussion regarding talc and ovarian cancer, by showing that, in some patients, talc can migrate to multiple organ sites in the pelvic area from perineal application. Recently, we showed that birefringent particles were detected by polarized light microscopy and shown to be talc by scanning electron microscopy / energy dispersive X-ray analysis (SEM/EDS) and/or Raman spectroscopy in surgical tissues from patients with ovarian carcinoma and perineal talc exposure. These sites included ovary, cervix, fallopian tube, uterine serosa, and pelvic lymph nodes.
Fibrous talc, with greater carcinogenic potential as determined by the International Agency for Research on Cancer, may be seen in pelvic tissues. Besides ascending through the reproductive tract, fibrous and non-fibrous (platy) talc, as well as other toxic materials such as asbestos, may enter pelvic organs by first accessing small submucosal lymphatics in the genital tract’s lower part.
Tissue-based analytic methods (including direct SEM/EDS examination of tissue sections cut onto PermanoxTM slides, and in situ SEM, where a paraffin block is examined directly by SEM under variable pressure partial vacuum) are critical in the detection of talc and asbestos, since they preserve tissue architecture, and show talc and/or fibrous particles within their histologic environment. Combined with correlative light microscopy, this enables one to see geographic landmarks and associated infiltrates, which helps inform exposure origin and pathophysiologic significance.
Stephan Goodman (Microscopy Innovations, LLC)
Automated and Very Fast Preparation of Biological Specimens for Transmission and 3D Volume Electron Microscopy
The preparation of biological specimens for TEM and 3D volume electron microscopy (3D-EM, including serial block face and FIB-SEM), is done by successively treating specimens with fluid reagents that include aldehyde and osmium tetroxide fixatives, heavy metal stains, dehydration solvents, and polymer resins. The typical process of manual delivery of these reagents requires 1-3 days for TEM, and nearly a week for 3D-EM preparation. This presentation will describe programmable automated preparation using the mPrep ASP-1000, that cuts the elapsed preparation time for TEM to just 1-3 hours, and cuts the preparation time for 3D-EM to ~1 day. Examples will include multiple mammalian tissues (liver, brain, heart), yeast, and flatworms that were prepared at several different universities and research institutes.
Jules A Gardener (Center for Nanoscale Systems, Harvard University)
Using a high-speed direct detection camera for ultrafast 4D-STEM data collection
Jules A Gardener1, Barnaby Levin2, Austin J Akey1, David C Bell1
1 Center for Nanoscale Systems, Harvard University
2 Direct Electron
The technique of 4D-STEM uses an electron sensitive pixelated camera to record STEM data, rather than traditional single-value-per-position bright field and dark field detectors. As a result, much more information about local distortions due to local crystal structure, defects, electric and magnetic fields can be probed. We will show our recent advances by using an ultrafast direct detection camera with high dynamic range at frame rates of 2000 frames per second, and that this is beneficial for acquiring ultra-high resolution STEM datasets with improved signal to noise and minimized scan/drift aftefacts.
