Exhibitor Tech Talks

Exhibitor Tech Talk Schedule a of 9/18/18

 

SUNDAY, DECEMBER 9, 2018MONDAY, DECEMBER 10, 2018TUESDAY, DECEMBER 11, 2018

 

8:15-9:15 am – Theater 1, Learning Center

Allen Institute for Cell Science

Cell data in the classroom: using the Allen Cell Explorer for high school and college education

Presenters: Carlos C. Goller, Thomas Martinez, Eric A. Shelden, Kaitlyn Casimo, Graham T. Johnson

Level: Introductory

The Allen Cell Explorer (allencell.org) is the Allen Institute for Cell Science data portal. It provides an unprecedented view into the organizational diversity of human stem cells, giving public access to large-scale 3D imaging data, predictive models, detailed methods, cell lines, and analysis tools. Three educators using various features of allencell.org in their classrooms will present their curricula and data applications. At the high school (AP Biology) level, we guide students through human cell image data on allencell.org that show variation in cell shape, structure, and dynamics. These lessons enhance the proteins and cellular pathways unit of the AP Biology curriculum with research-grade data and a view into the process of real-life science. At the introductory college level, we expose students to high-throughput approaches (HTA) and high-content microscopy datasets to engage them in the analysis of novel data. Students demonstrate applications of HTA via the analysis of image datasets from allencell.org during their transition from classroom-based learning to research positions. Advanced college/early graduate school students conduct virtual experiments using image-based cell data. In one virtual experiment, students analyze fluorescence microscopy images to categorize cells into cell cycle phases. They must then calculate the relative durations of the cell cycle phases while recommending experimental design parameters to maximize the accuracy of this approach.

 

 

8:15-9:15 am – Theater 2, Learning Center

Thermo Fisher Scientific

Innovations in Genome Editing Delivery and Transfection in Primary and Immune Cells

Presenter: Xavier de Mollerat de Jeu

Level: Intermediate

The talk will cover our latest transfection innovations for CRISPR genome editing, immune cell and in vivo delivery applications. Topics included in the talk: Nucleic acid delivery solutions for hard-to-transfect and primary & immune cells; Delivery of genome editing tools, including cas9 protein with electroporation or transfection reagents; High-titer production solutions for Lentivirus; In vivo delivery of mRNA and siRNA with Invitrogen™ Invivofectamine™ reagents.

 

 

9:30-10:30 am – Theater 1, Learning Center

Thermo Fisher Scientific

Using 3D organoid culture to assess disease aggressiveness in genetically engineered mouse models of prostate cancer

Presenter: Kristine M Wadosky, PhD

Level: Intermediate

Metastatic prostate cancer (PCa) patients who are resistant to targeted therapy can develop aggressive variants with neuroendocrine features. Our laboratory has published a genetically engineered mouse model (GEMM) that lacks Rb1 tumor suppressor and recapitulates this clinical course. Ezh2 histone methyltransferase is overexpressed in Rb1-deficient PCa and we made a new PCa GEMM that lacks Rb1 and Ezh2. Loss of Ezh2 increases PCa aggressiveness—unexpected since Ezh2 has been shown to be an oncogene. However, loss of one allele of Ezh2 appears to increase survival in Rb1-deficient PCa. These data indicate that Ezh2’s effect on PCa aggressiveness is dose-dependent. We have established PCa organoids from our original Rb1-deficient GEMM and those that lack one or both alleles of Ezh2. Using organoid imaging techniques, we are further investigating the differences in aggressiveness between our Rb1- and Ezh2-deficient PCa GEMMs. We are also using methods from neural cell culture to assess neuroendocrine characteristics of these PCa organoids and how Ezh2 expression affects these features. Finally, 3D organoid culture is allowing us to investigate molecular mechanisms driving PCa aggressiveness in the context of Ezh2 loss—a far less work-intensive approach than mouse genetics. This research can model the methods necessary to establish a 3D organoid culture system to simulate the tumor characteristics of GEMMs for any cancer type.

 

 

10:45-11: 45 am – Theater 1, Learning Center

Double Helix Optics

Double Helix Optics: Engineered PSF technology for 3D super-resolution light sheet and single molecule imaging and tracking

Presenter: Anurag Agrawal, PhD

Level: Introductory

Conventional 3D-light microscopy has allowed for keen insights into biological questions, but it is often limited in spatial resolution, imaging depth, or speed. Here, we introduce Double Helix Light Engineering™, which offers a library of engineered point-spread functions (PSFs) for a variety of extended depth 3D imaging applications. Our SPINDLE® optical module, along with this library of phase masks, is designed to seamlessly integrate with existing microscope platforms. This module currently enables nanometer-scale extended depth single molecule imaging and 4D particle tracking, and is compatible with illumination techniques including HiLo, TIRF, and Light Sheet. Further live cell imaging modalities are being developed, with the aim to extend imaging depths while reducing sample photo damage. Our 3DTRAX™ analytics software—designed as an easy to use ImageJ/Fiji plugin—has been optimized to provide detailed 3D structure and tracking information. This integrated platform is able to achieve average localization precision values below 20 nm laterally and below 25 nm axially with a 3-5x extension in depth range. We show 3D super-resolution reconstructions of subcellular structures, including actin, tubulin, stress granule cores and viral replication centers. In addition to these applications, we show how Double Helix Light Engineering can be used for extremely high-depth molecule and particle tracking with 7-10 times the depth of conventional methods.

 

 

10:45-11: 45 am – Theater 2, Learning Center

Thermo Fisher Scientific

Cells-to-CT™: A fast way to kill your cells for gene expression analysis

Presenter: Laura Chapman

Level: Introductory

You’ve taken painstaking measures to care for your cells, to keep them healthy and thriving. But now it’s time to crack them open and see what’s going on inside! Cultured cells are a powerful tool for in vitro analysis of gene expression.  Researchers use cells to screen gene modulation technologies or small molecule effects on gene expression and phenotypes. Cells are an excellent tool for screening and often lead to large experiments with many plates. Thus, a quick and easy solution is preferred, while maintaining sample integrity. At Thermo Fisher Scientific, we have a solution specifically for this: the Cells-to-CT™ product line. Cells-to-CT™ is very fast RNA preparation, 7 minutes, and user friendly, with only 2 pipetting steps done in the culture plate at room temperature. This “direct lysis” overcomes RNA loss by skipping the “purification,” and improves sensitivity with optimized reagents for lysis, RT and qPCR. The lysates go directly into qRT-PCR, providing gene expression data in 40 minutes for 1-Step or 90 minutes for 2-Step. In this Tech Talk, we will review gene expression applications for Cells-to-CT™ samples: siRNA transfection, single cell ESC differentiation, and targeted RNA Sequencing with AmpliSeq™ by NGS. The Cells-to-CT™ kits offer flexibility for low or high throughput sample preparation and are highly amenable to liquid handlers. Save your time and get your answers quicker, without diminishing your sample integrity with Cells-to-CT™.

 

 

12:00-12:45 pm – Theater 1, Learning Center

Bitplane -IMARIS

Analysis of 3D/4D Microscopy Images in Cell Biology – Imaris innovations advance the processing and analysis of complex datasets with a comprehensive and batchable environment

Presenter: Meredith Price – Product Manager, Imaris

Level: Intermediate

Recent releases of Imaris improved the performance when counting, tracking, and measuring morphological features of cells and other objects within large microscopy images. This Tech Talk will highlight 1) those Imaris innovations, 2) the newest Imaris family member, Imaris Stitcher, which aligns and stitches tiled images, and 3) the advances to be released in Imaris 9.3. Imaris 9.3 will provide a workflow for users who require automated image processing and analysis of their datasets within one comprehensive software environment. The features introduced in the new version are relevant to users segmenting cells, tissues, organs; tracking cells or organelles; or to those who are analyzing the relationship of one population of biological objects to another. We will illustrate Imaris’ unique, user-friendly tools for preparing a workflow complete with the comparison of results across groups of images.

 

 

12:00-12:45 pm – Theater 2, Learning Center

Carl Zeiss Microscopy

Celldiscoverer 7 and 3D-Cell Culture – High Resolution meets Screening; Structured illumination microscopy (SIM) with two-dimensional illumination patterns

Presenters: Dr. René Buschow, and Dr. Rainer Heintzmann

Level: Intermediate

Please join ZEISS for three informative talks discussing relevant topics from leaders in microscopy and cell biology. From 12:00-12:30 Dr. René Buschow will discuss using the imaging and analysis pipeline he and his team have developed to image 3D cell cultures using the ZEISS Celldiscoverer 7 automated imaging platform. From 12:30-1:00 pm Dr. Rainer Heintzmann will discuss theoretical and experimental results obtained from imaging live cells with two-dimensional structured illumination patterns. In our other Tech Talk from 1:00-1:30 Dr. Eric Betzig will discuss results obtained with a newly developed gentle, high speed, high resolution imaging technique called grazing incidence structured illumination (GI-SIM). For more detailed information, please visit www.zeiss.com/microscopy/ascb

 

 

1:00-1:45 pm – Theater 1, Learning Center

ACEA Biosciences, Inc.

Unbiased Functional Identification & Therapeutic Targeting of Tumor Neoantigens and Exploiting a Novel Cell Death Mechanism for Selective Killing of Cancer Cells Upregulating Homologous Recombination

Presenters: Dr. Stephen Schoenberger, La Jolla Institute, and Dr. Maria Soloveychik, SyntheX

Level: Intermediate

1) Tumor-specific mutations that are recognized by an individual patient’s T cells are called neoantigens, and can form the basis for personalized cancer immunotherapy. Neoantigens can now be readily identified through genomic sequencing and immune responses to them can be generated through vaccination and measured at the level of single cells. Thus, all the tools needed to evaluate and optimize personalized immunotherapeutic approaches to cancer treatment are in place.

2) At SyntheX, we have developed ToRPPIDO, a plug-and-play drug discovery platform for the identification of short peptide sequences and macrocycles that are capable of disrupting or bridging intracellular protein-protein interactions (PPIs) of interest in a cell-based system. Homology-directed DNA repair (HDR) plays a crucial role in maintaining genomic stability in cancer cells and is typically induced by oncogenes such as Myc, CycE, and KRas. In accordance, overexpression of HDR pathway components correlates with poor prognosis and chemo-resistance in most tumors, including currently untreatable pancreatic and biliary tract cancers. Application of our screening platform, ToRPPIDO, toward a crucial PPI within the HDR pathway led to the discovery of STX100. Subsequent derivatives of STX100 that are cell penetrant and proteolytically stable were tested using an xCELLigence instrument for selective activity against various cancer cell lines that overexpress HDR.

 

 

1:00-1:45 pm – Theater 2, Learning Center

Carl Zeiss Microscopy

Visualizing intracellular organelle and cytoskeletal interactions at nanoscale resolution on millisecond time scales

Presenter: Dr. Eric Betzig

Level: Intermediate

Please join ZEISS for three informative talks discussing relevant topics from leaders in microscopy and cell biology. From 12:00-12:30 Dr. René Buschow will discuss using the imaging and analysis pipeline he and his team have developed to image 3D cell cultures using the ZEISS Celldiscoverer 7 automated imaging platform. From 12:30-1:00 Dr. Rainer Heintzmann will discuss theoretical and experimental results obtained from imaging live cells with two-dimensional structured illumination patterns. Finally, in this Tech Talk from 1:00-1:30 Dr. Eric Betzig will discuss results obtained with a newly developed gentle, high speed, high resolution imaging technique called grazing incidence structured illumination (GI-SIM). The final 15 minutes will be Q&A. For more detailed information, please visit www.zeiss.com/microscopy/ascb

 

 

2:00-2:45 pm – Theater 1, Learning Center

Andor Technology

Dragonfly: Subcellular SRRF-Stream Super-resolution to Big Specimen Imaging with Imaris Stitcher

Presenter: Geraint Wilde, PhD – Product Manager, Andor

Level: Intermediate

Keywords: Confocal, TIRF, live cell, Developmental models, organoids, cleared tissue, 3D cultures. The Andor Dragonfly high-speed confocal has developed significantly in the last year. We have increased the number of imaging applications, and broadened the sample range that can be addressed with our confocal solution. This presentation will: 1) Introduce a new model of Dragonfly that can be used for applications that fit better with upright microscopes, e.g., deep imaging of cleared specimens, or use of dipping objectives; 2) Demonstrate the power of the newly embedded SRRF-Stream super-resolution technique perfect for live cell imaging (e.g., mitochondria, cytoskeleton) or fast SR imaging of fixed samples; 3) Demonstrate the ability to capture and stitch large 3D data at high-speed for large sample imaging. Come and see the high-speed, high-productivity, workflow Dragonfly offers from subcellular to whole specimen imaging. We can improve your time to publish by at least a factor of 10!

 

 

2:00-2:45 pm – Theater 2, Learning Center

Labviva

A New and Unique Approach for Relevant Scientific Information

Presenter: Siamak Baharloo, PhD

Level: TBD

One of the time-consuming activities in any research laboratory is the literature search for identification of the most appropriate reagents and instruments that will allow researchers to faithfully conduct their experimentation. Here we will introduce a methodology based on Natural Language Understanding (NLU) to organize, summarize, and understand large pools of scientific literature, protocols, and data sets with the aim of identifying research reagents that specifically meet the criteria defined by the researchers. Through the utilization of controlled ontologies, sentiment analysis, mined banks of terms/synonyms, and neural networks, chains of decision making can be structured that enable the technique, purpose, and outcome of a product to be determined in large batches of published material and then summarized for the researcher. Advancements in highly-scalable cloud infrastructure enable these techniques to be utilized more frequently for larger data sets, dramatically increasing the value of these techniques for the modern lab as queries against the output of these activities can be made in real-time, enabling its use in mobile devices embedded within the processes in the laboratory.

 

 

3:00-4:00 pm – Theater 1, Learning Center

ALVEOLE

Fine-tuning the mechanical and biochemical properties of in vitro microenvironments with a versatile and contactless photopatterning technology: PRIMO

Presenter: Pierre-Olivier Strale, PhD – Alveole Laurent Blanchoin, PhD – CytoMorpho Lab, CEA grenoble, CNRS

Level: Intermediate

In vivo, the cellular microenvironment has a crucial impact on the regulation of cell behavior and functions such as cellular differentiation, proliferation, and migration. One of the challenges confronting cell biologists is to mimic this microenvironment in vitro to more efficiently study living cells and model diseases. Here we present PRIMO®: a versatile, contactless, and maskless UV projection system, which allows researchers to engineer custom in vitro microenvironments and fine-tune their topography and biochemistry in a highly flexible and reproducible manner. We will first show that the projected UV light can be used to structure cell culture substrates via photopolymerization of: a) photosensitive resists to create molds onto which elastomeric solutions can be polymerized (microfabrication), b) and of the most commonly used hydrogels (microstructuration). Then we will show that PRIMO® is a suitable tool to print biomolecules on all cell culture substrates (including glass, plastic, soft or stiff substrates, textured surfaces, etc.) with an exquisite control over protein densities (micropatterning). Some of our users will then share their research conducted with PRIMO, in cell biology and microfluidics, from sub-cellular to cell population levels: reconstitution of the cytoskeleton with purified proteins, focal adhesions, cytoskeleton organization, force measurement, single cell confinement in 3D, cell migration, tissue engineering, spheroid formation, etc.

 

 

3:00-4:00 pm – Theater 2, Learning Center

Yokogawa Electric Corporation

Super Resolution Confocal Scanner Unit CSU-W1 Sora

Presenter: Takuya Azuma: Chief designer of CSU-W1 Sora, Yoshitaka Sekizawa: Product manager of CSU-W1 Sora

Level: Intermediate

Yokogawa will introduce our brand-new product “CSU-W1 SoRa.” This is a spinning disk based super resolution confocal scanner unit. In this talk, we will introduce features and principles of this product and we will show beautiful image samples taken by “CSU-W1 SoRa”. Features of “CSU-W1 SoRa”: 1) XY resolution of approx. 120nm. XY resolution has been improved by approximately 1.4x the optical limit based on spinning-disk confocal technology. Furthermore, a final resolution approximately twice the optical limit is realized through deconvolution. 2) Ideal for super-resolution live cell imaging. Just like the CSU, high-speed real time imaging can be performed with super-resolution. In addition, live cell imaging is possible, reducing bleaching and phototoxicity. 3) The CSU is easy to use. Super-resolution images can be observed in real time without any specific preparation of sample. Deep position observation is made possible through optical sectioning based on confocal technology. 4) Upgradable from CSU-W1. If you already have CSU-W1, you can add SoRa disk.

 

 

4:15-5:15 pm  – Theater 1, Learning Center

Bruker Corporation

Multiplexed quantitative single molecule localization microscopy with the Vutara 352

Presenter: Carl G. Ebeling, PhD – Worldwide Applications Scientist

Level: Intermediate

Single molecule localization microscopy has made a significant impact in the field of biology by increasing the resolving power of optical microscopy tenfold. One of the challenges within single molecule imaging has been the development and usage of adequate fluorophores for multicolor imaging. Advancing the scientific applications of single molecule imaging requires the ability to reliably image numerous biological targets within the same sample, visualizing the data, and performing quantitative analysis on the data set. Multiplexing methods, such as DNA-PAINT, Exchange-PAINT, and OligoSTORM remedy this by utilizing a fluorophore that is sequentially imaged, labeling specific biological targets during each imaging sequence. However, the optimized use of the above methods in a microscopy platform requires the use of an integrated fluidics system to allow delivery of the reagents to the sample over successive imaging sequences. To alleviate this challenge, Bruker’s Vutara 352 single molecule localization microscope has been designed to allow for multifaceted 3D single molecule imaging. Paired with Bruker’s custom fluidics module and control software SRX, the Vutara 352 integrates these multiplexing techniques into a single platform to allow for a systematic imaging modality. While SRX allows simultaneous imaging, localization and visualization, it also combines the numerous imaging sequences into a comprehensive localization data set. This allows for the interactive multicolor visualization of multiplexed data, and the integrated statistics module offers numerous computational tools to quantify the distribution, colocalization, and clustering of the various biological species targeted in the multiplexed data.

 

 

8:15-9:15 am – Theater 2, Learning Center

Leica Microsystems Inc.

Introducing an all new microscopic toolbox for functional imaging   

Presenter: Dr. Susanne Holzmeister

Level: Intermediate

The introduction of CRISPR/CAS has fundamentally changed the possibilities in biological manipulation. The creation of (fluorescently) tagged organisms or whole animal (knock-out) mutants is now feasible on a regular basis. To make use of these new possibilities, non-invasive observation is needed. Until now, only fluorescent microscopy was capable of fulfilling this requirement. A fundamentally new approach for functional imaging based on multiphoton microscopy and Fluorescence Lifetime Imaging will be provided. Learn how the utilization of spectrally tunable detection for multiphoton imaging available on the SP8 DIVE (Deep In Vivo Explorer) confocal platform combined with ultrafast fluorescent lifetime contrast of the SP8 FALCON (FAst Lifetime CONtrast) gives researchers easy access to the imaging of deep in vivo processes. The SP8 DIVE provides spectral capabilities that allows users to penetrate samples deeper and separates up to four spectrally different signals simultaneously, and a much larger number sequentially. The synergistic combination with FALCON provides additional information providing label-free imaging of key coenzymes like NADH, dye-separation of spectrally alike markers by their fluorescent lifetime or interaction of molecules using FLIM-FRET – deep within the tissue, in the most native context possible.

 

 

8:30-8:45 am  – Theater 1, Learning Center

Mizar Imaging

Bringing you the advantages of lightsheet for your live-cell fluorescence microscopy applications, now at high-/super-resolution

Presenter: Paul Maddox, PhD

Level: Intermediate

“Keep your cells happy and your objectives clean.” With that simple phrase Shinya Inoue captured an age-old dilemma of cell biologists: the light needed for optical microscopy also damages cells and bleaches fluorophores, creating particular problems for live-cell imaging. Structured planar illumination—lightsheet—reduces photo damage and bleaching. The Tilt lightsheet illumination system by Mizar Imaging offers these benefits of lightsheet, while also opening up for you the ability to use the objective lens of your choice, even high-NA, oil-immersion objectives. Designed to be an easy-to-use as well as high-performance system, the Mizar Tilt is an add-on unit that fits onto virtually any inverted microscope stand. Learn more about how the Tilt is being used for high- and super-resolution microscopy.

 

 

8:45-9:00 am – Theater 1, Learning Center

Fennik Life Sciences LLC

Introduction to Fennik Life Sciences & TheraKan™ System for 3D Cell Culture Analysis

Presenter: Lillian Cheng and Linda Cheng

Level:  Introductory

Fennik Life Sciences is a life science research company based in Kansas City and affiliated with the University of Kansas Medical Center. Its founders, Drs. Nikki Cheng and Wei Bin Fang, developed TheraKan, an innovative 3D culture device that creates a dynamic environment and closely models the complexity of living tissue and cell environments. They recognized the need for a more effective system after facing many challenges performing live cell imaging in mammary tissue in mice. The lab’s long-term goal is to understand how the immune system responds to breast cancer. Drs. Cheng and Fang realized the inherent difficulty of ensuring consistent tumor sizes as well as controlling for consistent numbers of immune cells recruited to the primary tumor. Their work using conventional methods required large number of animals resulting in high costs and intensive labor. Further, high-resolution multi-color images at thick areas of the tumor necessitated the use of specialized microscopy equipment, which was unavailable to them. Instead, they sought an alternative approach and developed the TheraKan™, which proved simpler and more cost-effective. The TheraKan™ utilizes a unique 2 chamber nested design with a simple swiveling mechanism allowing flow-through accessibility to cells and molecules from the outside. The device is designed for drug validation, biomarker expression analysis, cell migration, stem cell development therapies, immune cell recruitment, spheroid analysis, and more.

 

 

9:00-9:15 am – Theater 1, Learning Center

Double Helix Optics

Double Helix Optics: Engineered PSF technology for 3D super-resolution light sheet and single molecule imaging and tracking

Presenter: Anurag Agrawal, PhD

Level: Introductory

Conventional 3D-light microscopy has allowed for keen insights into biological questions, but it is often limited in spatial resolution, imaging depth, or speed. Double Helix Light EngineeringTM utilizes a library of engineered point spread functions that enable extended depth of field, for collection of more data with high precision axial information for 3D imaging and 4D tracking. Our SPINDLE® optical module, designed to be attached between most scientific microscopes and cameras, includes a library of phase masks to extend the imaging capabilities of microscopes for nanometer-scale single molecule imaging, light sheet, 3D particle tracking, and is currently being developed for additional live cell modalities. With this method, we are able to achieve average localization precision values below 20 nm laterally and below 25 nm axially with a 3-5x extension in depth range. We will demonstrate three implementations to enable extended depth nanoscale imaging: 1) extend the imaging capabilities of a TIRF microscope to nanometer scale 3D localization for super-resolution imaging and tracking; 2) use of Double Helix Light Engineering for extremely high-depth molecule and particle tracking with 7-10 times the depth of conventional methods; and 3) use of Double Helix Light Engineering in conjunction with Light Sheet microscopy to enable whole cell imaging and tracking at single molecule precision levels.

 

 

9:30-10:30 am – Theater 1, Learning Center

Cellecta, Inc.

Combining Cell Barcoding and CRISPR sgRNA Libraries with Targeted Gene Expression for Single Cell Genetic Analysis  

Presenter: Paul Diehl, PhD, COO, Cellecta

Level:  Intermediate

Pooled lentiviral libraries of CRISPR sgRNA to mediate genome-wide gene knockout have become an invaluable tool for uncovering the functional genetic drivers required for a biological response. Another type of pooled lentiviral library designed with unique DNA sequence tags have been used to label large populations of cells with unique cell-specific barcodes, which allows monitoring changes in sub-populations of cells with distinct phenotypes over time. Further, these barcode lentiviral libraries can be designed so that the unique barcode sequence is detectable in next-generation sequencing (NGS) RNA expression profiling assays (e.g., RNA-seq). As a result, investigators can analyze and link distinct molecular changes in sub-populations of cells with distinct gene expression profiles and phenotypes, such as drug resistance. The additional step, then, of incorporating these cell barcodes together with genetic effectors, such as CRISPR sgRNA libraries, makes it possible to tease out gene expression changes that result from specific genetic disruptions, and link these to the development of specific phenotypes. Cellecta will discuss work we are doing to develop this integrated platform that combines CRISPR sgRNA screens with single-cell genetic analysis.

 

 

9:30-10:30 am  -Theater 2, Learning Center

Bio-Rad Laboratories

The Best Practices and the New Best Fluorophores for the Best Western Blots

Presenter: Paul Liu, PhD, and Thomas Berkelman, PhD

Level: Intermediate

Generating publication-quality Western blots requires not just good technique but a thorough understanding of how each step in the workflow can affect data quality and reproducibility. New technologies are now available that make Western blotting more sensitive, quantitative, and reliable. In this talk we will discuss method optimization, technical best practices, and new products that improve Western blot detection and quantification. We will cover: How to prepare your cell/tissue lysate sample; How to estimate protein accurately; Gel/buffer chemistries – Choosing the right gel and buffer chemistries; Best practices for running a gel, transferring proteins to a membrane, and blocking; How to choose the best detection reagents; New products for fluorescent Western blot detection and quantification (StarBright™ Blue secondary antibodies and hFAB™ Rhodamine HKP primary antibodies); and Data analysis tools and tips. With this knowledge in your arsenal, you will run every Western blot more mindfully and the data you get will be more dependable and quantifiable. Our goal is to empower you to do better Western blots.

 

10:45-11: 45 am – Theater 1, Learning Center

GORYO Chemical, Inc.

Fluorescent Probes for Cell Biology

Presenter: Kenichi Maruyama

Level:  Intermediate

Fluorescent probes are an essential suite of reagents to probe and elucidate extracellular as well as intracellular processes in cell biology. A number of unique, highly specific reactive oxygen species (ROS) probes, metallo detectors, acid sensors and enzymatic fluors for glycobiology as well as proteases differentiated from existing probes through a single cleavage to liberate maximum fluorescence will be presented. A series of silicon rhodamine based fluors for super resolution live cell imaging under physiological conditions in microscopy, as well as the corresponding highly photostable fluorescent (Stella Fluor™) derivatives that have been adapted as labels for cell permeable as well as impermeable applications for imaging as well as flow cytometry applications will be described. Next generation calcium sensing and an innovative ratiometric probe to measure intracellular glutathione rapidly will be highlighted.

 

 

10:45-11: 45 am  -Theater 2, Learning Center

GE Healthcare

EDGE confocal: A new line scanning confocal technique for improved resolution and contrast

Presenter: Will Marshall

Level:  Intermediate

Microscopy is becoming a more important source of primary data in biological research. The ability to test multiple hypotheses in a single experiment through visualization and quantification of complex phenotypes (multiple markers, highly multivariate) facilitates more comprehensive studies in biologically relevant models. Due to recent advances in 3D cell culture and growing amounts of evidence that biological processes in these models are more representative of processes in vivo, there is a need for new imaging methods that improve image quality in thick samples without sacrificing speed or flexibility. We at GE have developed a new line scanning confocal modality that provides better background rejection, higher contrast, and improved z-resolution when compared with conventional point scanning or spinning disk modalities. The methodology is practical for wide scale adoption and amenable to both high resolution microscopy and high throughput automated imaging experiments in multi-well plates.

 

 

12:00-12:45 pm  – Theater 1, Learning Center

Andor Technology

NEW: Sona, a new back-illuminated sCMOS camera for the highest-sensitivity & largest field-of-view imaging— Protecting your specimen from photobleaching & toxicity; maximizing sampling & throughput

Presenter: Justin Cooper, PhD – Application Specialist, Andor

Level: Intermediate

Keywords: Live cell imaging, High content imaging, developmental biology, tissue imaging, organoids. Andor has just released the most sensitive back-illuminated sCMOS camera on the market. The flagship model, Sona 4.2B, presents an exclusive solution for capturing extremely large fields of cells or whole embryos with exceptional clarity. Great care has been taken to extend dynamic range and ensure linearity to ensure no valuable signal from your sample is lost, and that quantitative analysis is accurate. If you have an interest in capturing large volumes of data to improve your productivity, imaging large samples with as few fields as possible, or looking for a balanced solution between sensitivity, speed, and resolution for living samples, then join this presentation to find out more.

 

 

12:00-12:45 pm – Theater 2, Learning Center

Leica Microsystems Inc.

Introducing a fully integrated Cryo Electron Tomography workflow for your lab  

Presenter: Dr. Jan De Bock

Level: Intermediate

To fully investigate complex biological mechanisms, scientists require structural information about molecules within their subcellular context. To achieve this, the target molecules and their cellular environment need to be accurately resolved at subnanometer resolution. Leica Microsystems and Thermo Fisher Scientific have collaborated to create the first fully integrated Cryo Electron Tomography workflow that responds to these research needs and allows a fast screening of large areas and rapid determination of regions of interest in the electron microscope under cryo conditions. Safe and contamination-free sample and data transfer between instruments is ensured. Easy navigation to the cellular target regions via Correlative Light and Electron Microscopy (CLEM) is provided, being the basis for reliable results at subnanometer resolution.

 

 

1:00-1:45 pm – Theater 1, Learning Center

MilliporeSigma

Winning Westerns: Proven Strategies to Optimize Your Western Blots

Presenter: Natasha L. Pirman, PhD, Application Scientist, MilliporeSigma

Level: Introductory

Does Western blotting give you more trouble than expected? Do you feel like your precious samples are being wasted on bad Westerns? Join us and find out how you can improve your Western blots! In this seminar, you will learn general guidelines for performing and troubleshooting your Westerns, such as: • Choice of different blotting membranes • Parameters affecting blotting efficiency • Conditions for optimizing your immunodetection • Information on SNAP I.D.® 2.0: A faster way to perform immunodetection As the inventors of PVDF Immobilon® membranes, MilliporeSigma knows how informative a good Western can be. Bring your research questions to get the most out of this session.

 

 

1:00-1:45 pm – Theater 2, Learning Center

Synthego

CRISPR-Engineered Cells For Your Research  

Presenter: Kevin Holden, PhD – Head of Synthetic Biology at Synthego

Level: Introductory

The CRISPR-Cas9 technology is a powerful molecular biology tool that has revolutionized mammalian cell engineering. It is now relatively straightforward to generate gene knockouts, tag genes, or program specific genome changes such as single nucleotide variants in a wide range of cell lines and primary cells. Such engineered cells can be utilized for a variety of research purposes including drug discovery, disease modeling, gene therapies, and basic research. However, adapting this technology into the lab can require extensive optimization. Synthego has developed a software driven, automated platform for rapidly optimizing and generating CRISPR-Cas9 engineered cells for research purposes through a cloud biology solution. This allows scientists to focus more on their own research questions, and spend less time adapting and optimizing methods.

 

 

2:00-2:45 pm – Theater 1, Learning Center

Olympus America Inc.

The Olympus FV3000RS Laser Scanning Confocal Microscope: Speed and Sensitivity for Live Cell Imaging and Beyond

Presenter: Rebecca Murray, PhD, Product Manager, Confocal Microscopy Olympus America Inc.

Level: Intermediate

Olympus now offers the next generation FV3000RS laser scanning confocal microscope. Combining a large field of view resonant scanner and patented revolutionary spectral detection, the FV3000RS improves detection of dim signals at high speeds. Come learn about recent advances in confocal imaging technology as we discuss its applications for live cell imaging, thick tissue 3D imaging, and macro-to-micro imaging. We will also discuss the benefits of using red-shifted fluorophores for live cell imaging applications, such as reduced phototoxicity and improved tissue penetration. Olympus now offers the next generation FV3000RS laser scanning confocal microscope. Combining a large field of view resonant scanner and patented revolutionary spectral detection, the FV3000RS improves detection of dim signals at high speeds. Come learn about recent advances in confocal imaging technology as we discuss its applications for live cell imaging, thick tissue 3D imaging, and macro-to-micro imaging. We will also discuss the benefits of using red-shifted fluorophores for live cell imaging applications, such as reduced phototoxicity and improved tissue penetration.

 

 

2:00-2:45 pm – Theater 2, Learning Center

3i Intelligent Imaging Innovations

A spectrum of light sheet instruments optimized for different imaging demands

Presenter: Colin Monks, PhD

Level:  Intermediate

Light sheet technology has been at the core of many exciting recent developments in microscopy. These new instruments can image living cells at unprecedented speed with a minimal light dose. However, each instrument is optimized for only a subset of biological specimens. This talk will review the various light sheet architectures available and discuss the biological context where they are most useful.

 

 

3:00-4:00 pm – Theater 1, Learning Center

Horizon Discovery Ltd

Cell line engineering with CRISPR-Cas9 – tips and tricks to maximize success

Presenter: Vipat Raksakulthai , Field Application Specialist, Horizon Discovery

Level: Introductory

CRISPR-Cas9 is a versatile tool to discover more about your biological pathway or gene of interest. The simplicity of the CRISPR-Cas9 system has led to an explosion in applications, from single cell genomics to personalized medicine. In this Tech Talk, we will discuss key considerations when embarking on your gene engineering project to help minimize the bench time needed to reach your gene-engineering goals. Topics will range from designing guide RNA reagents to target your gene of interest to validating the resulting clones. We will also discuss the benefits of using an independent cell line model for confirmation of the phenotype. Key topics will include: Introduction to CRISPR-Cas9 genome engineering and Important considerations when initiating your gene engineering projects, including: Cell line optimization, Guide RNA selection, Donor design for knock-in studies, and Clone screening strategies and validation.

 

 

3:00-4:00 pm – Theater 2, Learning Center

Allen Institute for Cell Science

From images to information: new machine-learning based image processing toolbox for cellular organization

Presenter: Allen Institute for Cell Science

Level: Introductory

We will present the publicly available toolsets we have developed to tackle challenges in modern image-processing and analysis workflows. These include our new Python-based open source toolkit for streamlined 3D segmentation, which combines a set of traditional segmentation algorithms with an iterative deep learning workflow. This toolkit evolved from the challenge of developing successful 3D segmentation algorithms for a wide range (over 30) of intracellular structures in the Allen Cell Collection (www.allencell.org). We will demonstrate our straightforward workflow for applying this toolkit to your own 3D images of intracellular structures. We will also present a new method to get more information from brightfield imaging: our label-free method. It is designed to mitigate the challenges inherent in fluorescence microscopy. The method predicts 3D fluorescence directly from transmitted light images. It can be used to generate multi-structure, integrated images. Notably, training data for the method requires no manual annotation, little to no pre-processing, and relatively small numbers of paired examples, drastically reducing the barrier to entry. The methodology has wide potential use in many biological imaging fields and may reduce or even eliminate routine capture of some images in existing imaging and analysis pipelines, permitting similar throughput in an efficient, cost-effective manner.

 

 

4:15-5:15 pm – Theater 1, Learning Center

Nanolive SA

Label-free, long-term 3D analysis of organelles in living mammalian cells shows pre-mitotic organelle spinning: mitochondria, nucleus and lipid droplets in the spotlight

Presenter: Dr. Alexander Jones, Nanolive SA

Level: Introductory

Holo-tomographic microscopy (HTM) is a label-free non-phototoxic microscopy method reporting the fine changes of a cell’s refractive indexes (RI) in 3D. Cellular organelles such as lipid droplets and mitochondria which were dependent on chemical staining to be visualized, show a specific RI signature that distinguishes them with high resolution and contrast in HTM. Furthermore, thanks to the absence of phototoxicity proper of HTM, it is possible to follow the dynamics of mitochondria, lipid droplets as well as that of endocytic structures in live cells over long periods of time, even in the most sensitive type of cells (e.g., stem cells) which led us to observe, to our knowledge for the first time, a global organelle spinning occurring before mitosis1. Big announcements will follow the scientific talk. 1 Sandoz et al (2018), Label free 3D analysis of organelles in living cells by refractive index shows pre-mitotic organelle spinning in mammalian stem cells, Biorxiv, https://doi.org/10.1101/407239

 

 

4:15-5:15 pm – Theater 2, Learning Center

Thermo Fisher Scientific

Cryo-tomography: a new imaging technique for cell biology to peer at the inner workings of cells

Presenter: Elizabeth Villa

Level: Introductory

Studying the molecular machinery of cells from atomic detail to the cellular context and beyond is a great challenge for cell biology. This session will highlight how cryo -electron tomography allows researchers to peer inside cells and see proteins in situ—in their unperturbed functional environments—at high resolution and in 3D. The best possible structure preservation is guaranteed by sample vitrification, a freezing process so fast that it preserves structural integrity and functional interactions. Introducing innovative and user-friendly instrumentation makes this cutting-edge technology now more applicable for cell biology. This lecture will feature introductions into the Thermo Scientific Aquilos cryo-dual beam microscope and the tomography workflow utilizing Thermo Scientific’s cryo-transmission electron microscopes. The presentation will show how cryo-tomography is used to study the molecular architecture of the nuclear periphery, to understand Parkinson’s disease at the molecular level, and to peek into the inner life of bacteria.

 

 

5:30-6:30 pm – Theater 1, Learning Center

NanoSurface Biomedical, Inc.

NanoSurface Biomedical: Biomimetic Cell Culture Platforms for Enhancing Cell Biology Studies

Presenter: Nicholas A. Geisse, PhD

Level: Intermediate

Cells use structural and mechanical cues from the extracellular matrix (ECM) to regulate a broad spectrum of processes such as cell signaling, differentiation, division, and even life and death. Over the past few decades, the literature has demonstrated that many cell types cultured in conventional flat, rigid, and static culture conditions lack both structural and functional phenotypes seen in the body, and that the lack of extracellular cues contributes significantly to the disconnect between in vitro experimental results and in vivo observation. We will demonstrate that ECM-inspired substrate nanotopography drastically improves the structural and functional development of a variety of cell types. Specifically, we show how NanoSurface Cultureware and the NanoSurface Cytostretcher can be utilized to study the effects of cell-nanotopography interactions on adhesion, signaling, polarity, migration, and differentiation across many cell types and model systems including cancer biology, human epithelia, and cardiovascular function. Further, we will describe how the differentiation of stem cells can be enhanced by providing a more biomimetic culture environment, with particular focus on iPSC-derived cardiomyocytes. We will also illustrate how the combination of nanotopography and mechanical stretch can enhance the in vitro phenotypes of cells in culture.

 

 

5:30-6:30 pm – Theater 2,  Learning Center

Thermo Fisher Scientific

Advances in Image Processing Automation in Amira

Presenter: Trevor Lancon

Level: Introductory

Powerful image processing tools are essential to the modern microscopist’s workflow. Image sizes are growing as microscopes advance in quality and power, and with that the need for workflow automation is growing to be more important than ever. Amira rises to this challenge with the inclusion of recipes for image stack processing. Recipes are a mechanism by which tedious workflows of unlimited complexity are recorded, documented, and customized for application to further datasets. These recipes can be applied as a batch to 2D images comprising a stack, or as a sequence of 3D image processing operations. Saving a recipe as a simple text file allows the quick and easy transfer of expertise between colleagues or research sites via email. Results of recipes can include processed images, segmentation results, statistical measures, meshed surfaces and volumes, and more. During the talk, the recipe creation process will be explained and demonstrated for segmentation of a life sciences dataset. Follow creation, the recipe will be applied on further datasets. We’ll describe in detail how automation of this segmentation process eliminates the main bottleneck of the image processing workflow, and enables researchers to more quickly reach the results that push their research forward.

 

 

8:15-9:15 am – Theater 2, Learning Center

Leica Microsystems Inc.

NEW Leica THUNDER Imagers – Decode 3D biology in real time

Presenter: Oliver Schlicker, Product Application Manager Advanced Widefield Microscopy, Leica Microsystems

Level: Intermediate

Working in 3D biology with thick specimens such as organoids, spheroids, small animals, 3D cell cultures and tissue sections on a typical widefield microscope often leads to a loss of details caused by hazy images. In contrast widefield imaging is the perfect solution for combining highest speed with highest sensitivity in combination with lowest phototoxicity for physiological imaging. Leica Microsystems is proud to introduce its new THUNDER Imager—a family of widefield imaging solutions designed to deliver benchmark application performance in core life science applications. Leica Microsystems’ new THUNDER Imagers enable users to see through the haze using the latest opto-digital techniques using computational clearing to remove the typical haze inherent to all widefield images. THUNDER-powered solutions use minimally invasive widefield illumination without any additional mechanical complexities. Learn how these new imagers simplify your workflow, while allowing you to produce computationally cleared images at unprecedented speeds and quality.

 

9:30-10:30 am – Theater 1, Learning Center

Sapphire North America

ReZolve Scientific Photostable Fluorophores for live cell imaging by fluorescence microscopy and more

Presenter: Christie Bader, PhD

Level: Introductory

ReZolve Scientific’s range of fluorophores provide targeted insights on cell biology. The range of products are suited to confocal microscopy and include fluorescent probes that localize with polar lipids, mitochondria, and the endoplasmic reticulum. The probes use a unique metal core making them highly resistance to photobleaching, allowing for longer imaging with less signal loss. Applications include: • Cancer biology – ability to label polar lipids and lipid rich compartments that allows for easy comparisons and tracking of metabolic changes, which is important in cancer progression. • Neuroscience – detect lipid accumulation common to many neurological pathologies • Metabolic diseases – monitor mitochondria and changes in lipid content related to metabolic disease. Key advantages include: • Fast uptake of the dyes • Compatibility with other fluorophores, including GFP, allowing imaging of multiple attributes of the cell simultaneously • Use on live and fixed cells and tissue • Compatible with a range of fluorescent platforms and more. Unique properties: • Ability to label polar lipids • ER dye can wash-in wash-out for long term assays • Ability to detect mitochondria in fixed and frozen samples.

 

 

10:45-11: 45 am – Theater 1, Learning Center

Bruker Corporation

Advances in dye development and microscopy for live cell super resolution microscopy with the Vutara 352

Presenter: Robert Hobson, PhD – Applications Scientist

Level: Intermediate

Expanding the frontier of super-resolution imaging requires advances in both microscopy hardware and fluorescent labels. Here we describe a cooperative effort to improve both technological fronts with the ultimate goal of live-cell super-resolution microscopy. Bruker’s Vutara 352 super-resolution microscope has been designed for live-cell super-resolution microscopy with both high spatial and temporal resolution capabilities. The patented biplane module allows simultaneous two-color imaging in 3D while the sCMOS detector enables fast imaging of biological phenomena. Although this microscope system is capable of live-cell super-resolution imaging, it has been stymied by limitations in the current generation of live-cell-compatible fluorophores. Extant live-cell probes are either fluorescent proteins with low photon counts—and therefore low localization precision—or organic dyes, which require high laser power resulting in phototoxicity in living samples. To remedy this problem, we developed spontaneously blinking (SB) versions of the Janelia Fluor and Alexa Fluor dyes, which blink under physiological conditions at low laser power while still providing high photon counts. In particular, the spontaneously blinking Janelia Fluor 549 (SB-JF549) and red-shifted SB-JF646 are cell-permeable and are easily conjugated to HaloTag or SNAP-tag ligands, making them ready to use in live cell multi-color super-resolution experiments. The SB dyes, in combination with the Vutara 352, provide a powerful methodology for simultaneous imaging, localization and visualization of live-cell single-molecule localization data, while offering numerous statistical tools to quantify the data into publishable results.

 

 

10:45-11: 45 am – Theater 2, Learning Center

ChromoTek GmbH

One for All: Small Affinity-Tag & Nanobody for Multiple Capture & Detection Applications

Presenter: Dr. Klaus Herick

Level:  Advanced

We have developed a new epitope tag system based on a VHH, Nanobody®, or alpaca single-domain antibody. This VHH binds with high affinity to the Spot-Tag®, an engineered 12-aa sequence PDRVRAVSHWSS. Owing to the unique properties of the anti-Spot-VHH, the Spot-Tag capture and detection system is universally applicable. When covalently coupled to beads the anti-Spot Nanobody enables the immunoprecipitation and purification of Spot-Tag fusion proteins: • High affinity allows the purification of low-abundance proteins • Native elution possible with free Spot peptide • High chemical stability allows for extraordinary harsh buffer compositions and repeated use for purification Anti-Spot Nanobody conjugated to fluorophores allows the imaging of cellular proteins and structures using fluorescence microscopy: • Small size of Spot-Label leads to better tissue penetration • Spot-Label is the first detection tool directed against a small peptide tag that is ideal for super resolution microscopy owing to minimal label displacement. Examples for immunoprecipitation, Co-IP for MS analysis, affinity purification, immunofluorescence including super resolution microscopy, Western blot, ELISA, and CRISPR/Cas are given. The Spot-Tag system combines the high affinity and specificity of an antibody-epitope tag system with the stability and small size of an alpaca nanobody. This results in a universal tag-system that simplifies the purification and concurrent analysis of target proteins.

 

 

12:00-12:45 pm – Theater 1,Learning Center

MilliporeSigma

Dynamic Live Cell Imaging of Mammalian Cells Using CellASIC® ONIX2 Microfluidic Platform

Presenters: Cindy Chen, PhD, and Jun Park, PhD, Senior Scientists, MilliporeSigma

Level: Introductory

This workshop will cover the advantages of “dynamic live cell” imaging, where microenvironmental parameters such as flowrates, the perfusion of nutrients and reagents, and temperature and gas compositions can be precisely controlled on demand by software during the entire duration of a given imaging experiment. Overview of applications covering hypoxia, apoptosis, migration, and suspension immune cell imaging will be presented. Specific emphasis will be given to microfluidic designs targeted for use with different cell types as well as fluorescent probes for live cell imaging. Any scientists planning to start live cell imaging experiments, as well as experienced imaging scientists wanting to broaden their applications, will benefit from this workshop.

 

1:00-1:45 pm – Theater 1, Learning Center

Bruker Corporation

Cellular Imaging with Light-Sheet Fluorescence Microscopy: Ultra Gentle, High-resolution Imaging of Living Samples

Presenter: Dane Maxfield M.S., PhD – Sales Product Specialist

Level: Intermediate

Light-sheet fluorescence microscopy is a state-of-the-art imaging technique that allows for long-term 3D imaging at unprecedented speed across scales from single molecules to whole organisms. This presentation will focus on the Luxendo InVi SPIM, an inverted light-sheet geometry, which is optimized for long-term 3D imaging of live specimens with resolution better than on a confocal microscope and under meticulously adjustable conditions. We will describe our innovative sample mounting technique and demonstrate the ease of use for a variety of specimens, ranging from cell culture to organoids and embryos. Coupled with precise environmental control, the InVi SPIM allows for imaging of these samples for multiple days and for resolving subcellular structures without the photobleaching or phototoxicity that plagues standard imaging techniques like laser scanning or spinning disk confocal microscopy. You can expect to learn the advantages of light-sheet fluorescence microscopy for high resolution cellular imaging and how this technique can be adapted to a multitude of different samples.

 

 

2:00-2:45 pm – Theater 1, Learning Center

Bruker Corporation

Cell Mechanics with Atomic Force Microscopy (AFM): From Modulus Mapping to Measuring Cell-Surface Interactions

Presenter: Andrea Slade, PhD – BioAFM Product Manager

Level: Intermediate

Mechanobiology-related studies aimed at understanding how cells exert and respond to forces in their environment have become an important area of cell biology research. Examining the effects of forces on cells has a wide range of applications from understanding disease pathology to the development of tissue engineering devices. While operable in fluid environments under near-physiological conditions, atomic force microscopy (AFM) not only allows direct examination of the nanoscale structure of cell membrane surfaces but it also provides unique opportunities to quantitatively measure the nanomechanical properties of living cells and tissues. In addition, the integration of AFM with advanced light microscopy techniques (e.g., confocal, super-resolution, etc.) enables direct correlation of these mechanical properties with fluorescence imaging datasets. Please join us for this informative seminar where we will introduce our complete family of BioAFMs, including the latest JPK systems to join Bruker. We will describe the most recent advances in Bruker’s BioAFM technology, focusing on various examples of how our industry-leading capabilities are enabling new possibilities for novel cell mechanics studies, both in real time and in situ.

 

 

3:00-4:00 pm – Theater 1, Learning Center

MilliporeSigma

Duolink® PLA: A Powerful Tool to Study Protein-Protein Interactions and Signaling Pathways

Presenter: Tracy Adair-Kirk, PhD, Principal Scientist, MilliporeSigma

Level: Introductory

The ability to study concerted movements, modifications, and interactions of proteins within a cell in a multiplexed fashion is key to unraveling the fundamental mechanisms of biology and disease states. However, the low-level expression of many proteins, combined with the transient nature of their interactions, makes analyzing these processes quite difficult. Duolink® in situ proximity ligation assay (PLA) offers a solution to overcome these problems. Duolink® PLA is both highly selective and sensitive, resulting from dual antibody recognition and rolling-circle amplification, which occurs only when the two PLA probes are in close proximity. Protein targets can be readily detected, quantified, and localized with single molecule resolution in unmodified cells. Duolink® PLA can be adapted for use on suspension or adherent cells, tissue sections, and multiwell plates, making it an ideal method for performing high-throughput screening of drugs, inhibitors, or monoclonal antibodies, target validation, and disease pathway analysis. In addition, Duolink® flowPLA now allows the detection of very low abundant proteins and protein interactions by flow cytometry. Furthermore, Multicolor Duolink® PLA allows multiplex detection of up to 4 protein events (e.g., protein interactions or modifications) within a single assay. These recent advances in the Duolink® PLA technology will allow researchers to generate more robust data in fewer tissue or cell samples.