This year, at ASCB2015, Nanolive (http://nanolive.ch, booth #1322) will launch its revolutionary microscope, the 3D Cell Explorer which images living cells instantly, in 3D and 4D (http://goo.gl/mtcm7o), without labels at a resolution below the diffraction limit of light (200nm; http://nanolive.ch/3d-cell-explorer/).
The refractive index distribution within the cell is measured at each pixel and the researcher can decide which parts of the cell to visualize by digitally staining them in contrasting colors, without interfering with the cell’s normal physiology (http://nanolive.ch/technology/). Join our Tech Talk on Sunday at 9:30AM (Theater 1) and don’t miss our launch event on Monday 5:30-7:45PM in the Theater 2, Learning Center (https://www.ascb.org/2015meeting/tech-launches/)
Presenter: Dr. Christian Klose
Complex lipid compositions of a variety of samples, like tissues, cells, cellular preparations, organelles, body fluids and others are challenging to analyze quantitatively in a single, straightforward measurement.
Here we present a fully quantitative lipidomics technology characterized by high precision not only on different days but also in different laboratories. At the same time our technology allows for high lipid coverage, without compromising the throughput.
To scrutinize the feasibility of our approach we applied it to human blood plasma, where it proved:
- to be comprehensive, covering 22 different lipid classes encompassing more than 200 lipid species;
- to be high-throughput amenable, allowing for the analysis of hundreds samples per day;
- to achieve absolute quantification of individual lipid molecules, by inclusion of lipid class-specific internal standards
Presenter: Mark A. Tsuchida
µManager (Micro-Manager) is free and open source software for optical microscope control and image acquisition. µManager allows you to run imaging experiments using a wide range of devices including major microscope stands, scientific cameras, stages, illuminators, and more, all through a simple and intuitive interface. In this short presentation, we will introduce you to µManager’s features and capabilities and provide a glimpse into µManager’s extensibility and customizability. We will also discuss why you should care about open source software for science and introduce the µManager user support services available from Open Imaging.
Presenter: Dr. Randall K. Wetzel, PhD
Fluorescently-labeled antibodies and cellular dyes can be combined with simple cellular analysis devices to quickly and easily monitor biological processes. These include cell health, viability, immune or other cellular signaling, protein expression, metabolism, cell cycle, apoptosis, and toxicity in multiplex whole cell or lysate-based sandwich assays. In this presentation, we will review common cellular assays and describe simple protocols and reagents to enable these assays.
Presenter: Xavier de Mollerat du Jeu, PhD Director R&D Life Science at Thermo Fisher Scientific (Carlsbad, USA)
Xavier de Mollerat du Jeu is leading the R&D efforts for the development of new delivery solutions of Nucleic acids, including Lipofectamine® and Invivofectamine® product lines
During this seminar he will focus on the latest advances in:
- Nucleic acid delivery solutions for hard-to-transfect and Primary cells
- Delivery of Genome editing tools- including cas9 protein
- High titer Lentiviral production solutions
- In vivo delivery of RNAi and mRNA using invivofectamine3.0
Presenter: Matthew H. Cato, Applications Scientist
Tools to analyze single cells for their discrete characteristics are becoming increasingly mainstream, bringing with it significant scientific momentum. Scientists understand that cell populations are heterogeneous and bulk measurements can mask events when using the law of averages. However, current genomics approaches introduce variables that can result in over/under representation of certain cell types. Ultra-sensitive fluorescent in situ hybridization (FISH) that relies on signal amplification rather than transcript amplification provides an outstanding single-cell validation tool. These techniques illustrate subcellular, population dynamics/cell frequencies, or indicate morphological context. ViewRNA® and PrimeFlow® FISH platforms for microscopy and flow cytometry incorporate a proprietary probe design using branched DNA (bDNA) signal amplification technologies that result in excellent specificity, low background, and high signal-to-noise ratios.
Presenter: Dr. Kamala Tyagarajan, Ph.D
Understanding the linkages between cell cycle and cell death pathways has become increasingly important to elucidating the action of toxins and anti-cancer compounds, as well as the fundamental mechanisms of cell division. Cell biology research therefore requires simple, accessible tools to determine impacts on both cell cycle and cell health in order to characterize the pathways by which cells are affected by treatments or conditions. We will present simplified flow cytometric methods using the Muse© Cell Analyzer to study cell cycle distribution together with data from well-known apoptosis and cell death assays. You will learn how the parallel study of cell cycle and cell stress/death can provide enriched information on the impact of treatments, and provide a comprehensive picture of cellular status and health.
Presenter: Bob McCarthy
Crude collagenase is commonly used to recover cells from tissue. The benefit of using this low cost product comes at a price: the inability to define the biochemical components responsible for the success of cell recovery. As life science research transitions toward translational medicine, these reagents should be better defined. This tech talk provides an overview of how collagenase enzymes are manufactured, the limitations of using crude enzymes in cell isolation procedures, and the advantages of using a new defined, enriched collagenase product (DE Collagenase) that contains primarily collagenase and a purified protease. Comparison of the biochemical characteristics of crude and DE Collagenase enzymes will show how definition of key enzymes responsible for cell isolation minimizes the need to prequalify lots prior to purchase.
Presenter: Teng-Leong Chew, PhD Director, Advanced Imaging Center Howard Hughes Medical Institute Janelia Research Campus
Visualizing and understanding complex biological processes demands the integrated efforts of biologists and physicists. The mission of the Advanced Imaging Center (AIC) is to make cutting-edge imaging technologies developed at Janelia widely accessible, and at no cost, to scientists, years before they become commercially available. This unique imaging center is thus uniquely positioned to empower investigators with tools currently not widely available elsewhere, such as the lattice light sheet microscope recently developed by Dr. Eric Betzig. In alignment with Janelia’s philosophy of encouraging bold and risky science, the AIC welcomes high-risk-high-gain projects that may challenge the current paradigm and provides full through Janelia’s in-house imaging experts and research infrastructure. This seminar will present the technical capabilities and the application process of the AIC.
Presenter: Joseph Huff, Product Marketing Manager, Laser Scanning & Superresolution Microscopy
Learn about two new detection and acquisition strategies for the ZEISS Airyscan detection module for laser scanning microscopy. ZEISS Airyscan, a new detector concept designed for improved laser scanning confocal microscopy, enables the simultaneous increase of both resolution and signal-to-noise ratio over traditional confocal imaging. Both modes – sensitivity and two-photon – extend the Airyscan benefits of resolution and signal-to-noise ratio to address more sample types.
Presenter: Dr. Xufeng Wu, NHLBI NIH
Confocal microcopy is and has been a mainstay of light microcopy for decades given the flexibility of a commercially available system. However, the resolution of a confocal microscope is still diffraction limited with a lateral resolution of ~250nm. With the recent advent of superresolution techniques (PALM, STORM, STED, and SIM), new commercially available systems are being used to image beyond the diffraction limit. As a result, the increased spatial resolution novel information regarding cell structure and function can be obtained. Recently the new Zeiss Airyscan detector was introduced as another method of superresolution and represents a good blend of resolution, sensitivity and speed. In this session, Dr.Xufeng Wu will present her recent experiences using the Airyscan for imaging cyoskeletal dynamics, organelle, membrane trafficking, and three-dimensional.
Presenter: Manasa Gudheti, Ph.D.
Super-resolution microscopy has made a significant impact in the field of biological imaging. Most imaging has been targeted at fixed specimens with a few live-cell applications. The Vutara 352 super-resolution microscope has been engineered for live-cell applications through the optimization of spatial and temporal resolution in single-molecule localization imaging. Its sCMOS detector enables video rate imaging along with two color simultaneous imaging in live cell and 3D particle tracking experiments. The biplane based detection path enables imaging thicker samples such as whole mount Drosophila and offers deeper penetration into tissues.
The Vutara 352 also includes real time localization along with several statistical and live cell analysis features for processing data. In summary, the Vutara 352 microscope is a powerful super-resolution imaging and analysis tool.
Presenter: Melike Lakadamyall, Ph.D.
Super-resolution microscopy has emerged as a powerful tool to image cells at the nanoscale, giving important insights into biological processes at the molecular level. The revolutionizing impact of these methods has recently been recognized by the Nobel Prize/Chemistry. While exciting developments have significantly advanced the capabilities of super-resolution microscopy, important challenges remain in pursuing its applications in biology. One limitation is the compromise between spatial and temporal resolution that make these methods poorly suited to study dynamic processes. Another challenge is our limited capability to extract quantitative information from the super-resolution images, which is confounded by the photophysics of the fluorophores. I will show how we are developing new approaches to overcome these challenges and demonstrate novel biological applications of super-resolution.
Presenter: Luciano Lucas
Low-toxicity imaging technologies have enabled an expanding group of researchers to examine cellular processes over long periods and at sub-cellular resolution. Developmental, stem cell and cancer cell biology have all benefited greatly by employing techniques such as spinning disk confocal, light sheet and structured illumination microscopy. A major bottleneck felt by scientists is the efficient visualization and analysis of long 3D time-lapses. Building on the powerful file handling of Imaris and functionality of ImarisTrack and ImarisCell we are introducing a new tool set which enables researches to study cell division, track and interactively explore cell lineages. This talk will be the launch of Imaris 8.2. I8.2 tackles the bottleneck mentioned above thus enabling the scientific community to post and test previously impossible to test hypothesis.
Guntram Bauer, Carmen Gervais (HFSP), Carmen Gervais (HFSP)
Dan Kiehart (Chair HFSP CDA review committee)
Claire Walczak (former HFSP reviewer for research grants)
HFSP promotes international collaboration in the spirit of science without borders. Novel, innovative and interdisciplinary approaches to basic research are invited, with a focus on scientists who are early in their careers. This talk will present the funding programs and explain the common pitfalls that make applications unsuccessful and some practical advice on how to increase your odds of success. The program is suited for those interested in pursuing high quality, international research exchanges. With a focus on scientists changing fields or areas of study, it’s a unique program that fosters scientific independence and sets the stage for research careers at the frontier. The presenters will talk about HFSP opportunities, best practices in career planning, and provide tips on how to structure a competitive application.
Presenter: Dr. Daniella Steel
Genome engineering has been revolutionized recently by the emergence of CRISPR-Cas9, which has democratized the generation of custom engineered cell lines. Unfortunately, as many users are discovering, the reality isn’t always as simple as hoped.
Horizon is at the forefront of gene editing and has amassed the world’s largest collection of off the shelf or on-demand genomically modified cell lines (>20,000 X-MAN® models) available to support research programs worldwide. We will cover CRISPR-Cas9 as a tool for cell line generation, will discuss how to plan a CRISPR genome editing experiment to maximize your chances of success, and will focus on the applications of these cell lines, exploring through case studies the genomic modifications available to cell biologists, including knockouts, knockins, translocations and endogenous tagging.
Innovation in advanced imaging is a driving force for science. Leica Microsystems’ newest inverted microscope platform, the Leica DMi8, features modularity that allows users to build a powerful imaging system specific to your research and budget today without compromising essential functionality for the future. Examples of the platforms modularity include FRAP, confocal imaging, digital light sheet, and super-resolution STED and GSD.
Powered by its modularity, unique Infinity Port™ and next generation 19mm FOV imaging ports, the DMi8 allows users to see more of their data.
Presenter: Vratislav Kostal, Ph.D.
Digital holographic microscopy (DHM) is an emerging microscopic technique for high resolution, label-free observations of living cells. Compared to standard microscopy, DHM has the unique ability to record complete information about the light waves including intensity, DIC contrast and especially the phase shift. The quantitative phase imaging (QPI) provides unique insight into the distribution of cellular mass. Here, we present the latest version of our QPI microscope (Tescan QPHASE), which is designed specifically for long term studies of cellular dynamics and morphology. A unique illumination setup allows imaging of cellular mass and morphology with extreme axial sensitivity and no halo effects. The unique capabilities of the system are demonstrated by time lapse studies of cellular death, cancer cell proliferation and cell behavior in 3D environments.
Presenter: James Goldmeyer, Ph.D, Global Product Manager
The emergence of the CRISPR-Cas9 system to generate alterations in the mammalian genome has expanded the scope of functional biology and many parallels have been drawn between the CRISPR-Cas9 system and the RNAi pathway. While fundamentally different, the challenges of off-targeting and poor functionality can be inherent to both systems and cause concerns about data interpretation and reliability. Used together, these technologies can provide a robust insight into the functions of genes in mammalian cells. In this talk we will describe a novel approach to understanding the requirements of both function and specificity of crRNAs and present data supporting the development of the next generation of shRNA constructs.
The Opterra imaging system is a high speed confocal system designed for live cell, volumetric microscopy. The Opterra II builds upon the success of the first generation Opterra’s Swept Field Confocal (SFC) technology by offering rapid filter switching, quality wide-field/DIC imaging via bypass mode, enhanced optics for field uniformity, and sophisticated photo-stimulation capability. Flexibility in choosing various pinhole and slit apertures has allowed researchers to balance spatial resolution, speed and fluorescence intensity for each experiment.
Come learn about the variety of research enabled by the instrument including studies of wound-healing processes in the Xenopus laevis, neural development in zebrafish, tracking of multi-labeled cilia and others.
Presenter: Andrew Moore, Editor-in-Chief
Attend this Open Access workshop with Andrew Moore, a highly-experienced Wiley Editor. Andrew will be talking about Open Access; the changes it’s brought to the field, what you need to know, and what it means for you. Come and improve your understanding of Open Access and take the opportunity to ask Andrew any questions you have on the topic
Presenters: Alex Zambon, Chris Shumate
If a picture is worth a thousand words, then a movie should be worth tens of thousands of data points. The timely integration of affordable time-lapse live-cell microscopy provided by Etaluma, along with genetically encoded biosensors and quantitative image analysis approaches, have enabled our laboratory to develop a robust and extremely low cost screen.
Our genetically encoded biosensor harnesses cell-cycle-dependent transcriptional and post-transcriptional regulation of fluorescent reporter proteins that ultimately enable multi-parameter quantification of each of the main phases of the cell cycle (i.e. G0, G1, S, G2/M) at single cell resolution in living cells and tissue. This economical system is being applied to novel drug and gene discovery in regenerative medicine and cancer.
Presenter: Leanna Ferrand, Product Support Leader
Live cell imaging has long been one of the most challenging yet most rewarding applications in fluorescence microscopy. Recently, super resolution microscopy has allowed researchers to investigate previously unresolvable structural details in their samples. Combining these techniques to perform live cell super-resolution imaging creates additional challenges, but greatly increases the potential scientific reward. Advances in Structured Illumination Microscopy (SIM) have made biologically relevant live cell SIM a reality. SIM offers researchers a two-fold increase in resolution both laterally and axially, revealing those structural details previously unresolved with conventional microscopy. SIM requires the least amount of input light, thus reducing photobleaching and phototoxicity. SIM is compatible with standard fluorophores and sample preparation techniques, requiring minimal sample optimization and is becoming the most approachable live cell super resolution method.
Presenter: Bob Balderas, VP of Biological Sciences
Studying multiple aspects of a cell leads to a deeper understanding of cell biology, including reporter gene expression, cell proliferation and death, cell signaling, and genomics. We will review flow cytometry for cell biology, and discuss its evolution to allow simultaneous analysis of multiple parameters. Experiments performed on the BD Accuri™ C6 flow cytometer, followed by higher parameter assays run on a BD FACSCelesta™ flow cytometer, will be shown. Single cell gene expression studies using the BD FACSseq™ cell sorter with BD™ Precise Assays will be discussed. Topics include:
- Fluorescent protein analysis
- Multicolor panels
- Analysis of phosphorylated proteins at the single-cell level
- Rapid, easy cell isolation for genomic analysis
Presenter:Ross Whittaker PhD, MBA Product Manager
CRISPR-Cas9 systems provide a platform for high efficiency genome editing that can lead to innovative applications in cell engineering. However, the delivery of Cas9 and synthesis of guide RNA (gRNA) remains two steps that limit overall efficiency and general ease of use. In this presentation we will discuss the rate limiting steps in current CRISPR/CAS9 workflows and introduce streamlined methods to reduce the editing workflow timeline to just 4 days. Additionally, we will discuss our recently launched GeneArt Engineered Cell Models collection which can provide instant access to human cell lines containing Knockout and Knock-in mutations. This resource along with the streamlined workflows allow the researcher to the rapidly test their hypotheses and accelerate their research programs..
Presenter: Nick Dolman PhD, Sr. Staff Scientist, R&D, Thermo Fisher Scientific
The combination of light microscopy and fluorescent reporters offers an unparalleled view into the function of intact cells. Recently, the scientific community has witnessed many major innovations in fluorescence microscopy that have paved the way for exciting new discoveries. Invitrogen™ Molecular Probes™ reagents have represented a significant aspect of these key innovations, driving cutting edge fluorescent reporter development for four decades. In this seminar, we will highlight the breadth and depth of fluorescent probes available to the cell biologist of today, drawing on specific techniques as diverse as flow cytometry , single molecule and in vivo imaging to high content and multicolor time lapse imaging. Specific examples of assays to visualize key aspects of cell biology will also be described that include cytoskeletal dynamics, cell tracing, endocytosis, proliferation, autophagy, hypoxia, oxidative stress, and apoptosis. Furthermore, we will provide an overview of new probes and current constraints in serving emerging areas such as 3-D cellular models and super resolution microscopy.
Presenter: Steve Offer
Steve Offer, Assistant Professor of Pharmacology at the Mayo Clinic School of Medicine, describes the cellular metastasis models his lab has developed to better understand the molecular changes that occur during colorectal cancer metastasis. Steve will discuss the correlation between changes in the epigenetic landscape, microRNA expression, and gene expression, and how these changes affect tumor progression. Along with traditional approaches to study metastasis, Steve’s lab has leveraged the throughput and sensitivity of the xCELLigence real-time cell analyzer and the NovoCyte flow cytometer to test specific parameters of candidate metastasis-associated signatures identified by his lab. These results are expected to help identify early-stage indicators of later tumor aggressiveness and to facilitate the development of therapeutics designed to specifically target cancers based on discrete molecular profiles.
Presenter: Paul Diehl PhD.
The CRISPR/Cas9 system is a targeted gene-editing tool adapted from Streptococcus pygogenes and can be used to permanently knock out target genes. It has revolutionized the genomics research along with the earlier tool of RNAi. Genome-wide loss-of-function pooled screens provide a direct approach to identify genes regulating biological responses and find new therapeutic targets. While RNAi screens have proven effective, CRISPR/Cas9 provides an alternative screening approach. To complement our shRNA screening platform, we developed pooled sgRNA libraries for functional CRISPR knockout screens and compared results from sgRNA and shRNA library screens of PDX-derived cell lines. This talk will provide an overview of the functional genomics screening approaches and focus on comparison of the two available methods.
Presenters: Ivona Strug, Ph.D., Sara Gutierrez
Did you know a reliable Western blot of cell lysates starts before you load your sample on a gel? Learn how cell type, health, cell count, total protein concentration, and lysis buffer composition can affect your semiquantitative Western blot. You’ll learn how to:
- Obtain signals that are detectable but not supersaturated
- Perform cell lysis/extraction to ensure sample-to-sample consistency
- Choose a transfer membrane for excellent protein retention and signal-to-noise ratios
- Use IR spectrometry to obtain more reliable quantitation of total protein in cell lysates compared to Bradford/BCA assays
- Use IR spectrometry to improve separation and quality of SDS-PAGE
As the inventors of PVDF membrane, EMD Millipore knows how informative a good Western can be. Bring your research questions to get the most out of this session!
Presenters: Pierre-Olivier Strale, Ammar Azioune, Ghislain Bugnicourt, Yohan Lecomte, Makhlad Chahid and Vincent Studer
We describe how to perform high resolution multi protein micro-patterning using Light Induced Molecular Adsorption (LIMA). LIMA is based on a photo-initiator able to reverse the antifouling property of polymer brushes when exposed to UV light.
The density of adsorbed molecules scales linearly with the dose of UV and the low background allows for the sequential printing of multiple proteins. Our optical set-up (widefield DMD based projection system + epifluorescence microscope) allows us to generate patterns ranging from 500nm to 1 mm and controlled gradients of arbitrary shape. The range of application extends from the single molecule up to the multicellular scale with an exquisite control over protein density.
Altogether, LIMA allows for fast high resolution patterning of multiple proteins with applications to biomedical research.
Presenter: Paul Held
Image-based analysis is a powerful tool to assess cellular processes. In this workshop we will demonstrate how the Cytation 5 Cell Imaging MultiMode Reader can be used for live cell kinetic experiments involving:
- Gene-specific mRNA expression using SmartFlare probes
- Receptor activation through the use of genetically encoded biosensors that detect GPCR second messengers.
- Miniaturized RNAi knockdown experiments using microarray spotting technology
Other applications, such as cell cycle, viability, and migration, conducted in either 2D or 3D spheroids will be discussed.
Presenter: Elliot Fisher, Sangwook Choi
NanoSurface Biomedical’s ANFS (Anisotropically Nano-Fabricated Substratum) technology integrates mechanical characteristics of the in vivo cellular micro-environment into a familiar in vitro platform for daily use by researchers. Featuring topographical resemblance to the native extracellular matrix, NanoSurface Biomedical’s nanopatterned substrates provide a topographical and mechanical niche for cells, facilitating growth as if in vivo. High resolution microscopic images of cells cultured in ANFS dishes are made possible through the use of low auto-fluorescence surface materials; these products are ideal for confocal microscopy, phase contrast microscopy, live cell imaging and micro-manipulations. Currently available in 35mm culture dishes, 6-well plates, and 24-well plates.
Presenter: Anne Marie Quinn
G protein coupled receptors produce a myriad of effects through a variety of effectors. To study these effects, we created genetically-encoded fluorescent biosensors for cAMP and DAG. These sensors are analogous to the GCaMP and GECO Ca2+ sensors, in that the analyte binding domains are fused to a single fluorescent protein. A major advantage of single fluorescent protein sensors is that they can be combined for simultaneous measurement of multiple analytes. These sensors are robust enough for detection with epifluoresence microscopy or fluorescence plate readers. The sensors are packaged in viruses for delivery to mammalian cells. Changes in fluorescence are reproducible from cell to cell. cADDIs, a cAMP sensor detects both Gs and Gi-coupled responses. Upward and Downward DAG sensors detect Gq signaling.
Presenter: Sean Yu
VitaScientific is a life science supplier based on Maryland, USA. We constantly strive to bring the highest quality, most innovative products from around the globe to biomedical researchers in their quest to better the human condition. We pride ourselves on our customer service, technical support, and dedication to ensuring your experiment is a success.
We would like to introduce:
- A new electroporation technology that ensures uniformly high efficiency;
- A transfection product line that routinely outperforms the Lipofectamines;
- The only commercially CLARITY tissue clearing system for cutting edge 3-D imaging;
- Dedicated semi-automated Western Blotting imager, digital fluorescence microscope with high quality imaging sensors;
Visit us at www.VitaScientific.com for more innovative research tools for your lab.
Presenters: Dr. Gary Rondeau from Applied Scientific Instrumentation Inc. (ASI), Dr. Larry Shi & Dr. Dan Christensen from TOPTICA Photonics, Inc.
Dr. Gary Rondeau from Applied Scientific Instrumentation Inc. (ASI) will discuss the operation & advantages of the Dual Inverted Selective Plane Microscopy (diSPIM) system that was developed in collaboration with Dr. Hari Shroff and his group at the NIH/NIBIB. Recent developments will be discussed, which include operation modes for stage scanning, multi-color acquisition and auto-focus during long acquisitions.
Dr. Larry Shi & Dr. Dan Christensen from TOPTICA Photonics, Inc. will discuss features and benefits of TOPTICA’s broad portfolio of multi laser engine (MLE) products, specifically how it improves light sheet microscopy in the diSPIM system. Equipped with a proprietary, fully automated laser alignment system (COOL AC), the TOPTICA MLE is the only commercially available, turn-key laser light engine that guarantees consistent optical power over time.
Presenter: Yann Cotte
Nanolive (http://nanolive.ch, booth #1322) is launching the 3D Cell Explorer, its revolutionary microscope able to image living cells instantly, in 3D and 4D (http://nanolive.ch/applications/), without labels at a resolution below the diffraction limit of light (200nm). The refractive index distribution within the cell is measured at each pixel and the researcher can decide which parts of the cell to visualize by digitally staining them in contrasting colors, without interfering with the cell’s normal physiology (http://nanolive.ch/software/).
An engaging keynote from Nanolive’s CEO, Yann Cotte will be followed by a reception open to all ASCB participants (https://www.ascb.org/2015meeting/tech-launches/).
Presenter: Jennifer Welser-Alves
For over a decade, ScienCell Research Laboratories has helped researchers with their cell culture experiments by providing expert advice. We will discuss primary cell culture optimization techniques and why primary cells are necessary for validation of cell line studies and in vivo experiments. In addition, we will examine the importance of selecting the appropriate pluripotent stem cell expansion and differentiation media and illustrate the advantages of using pluripotent stem cells for your research. Lastly, learn how to advance your primary cell research by using our new GeneQuery™ qPCR Arrays kits for gene expression profiling.
Presenter: Daniel Sanchez and Vincent Schoonderwoert
Microscopy images suffer from artifacts like blurring, noise, and chromatic aberration which all distort your image and affect the ‘true’ object distribution. Fortunately, the most important distortion factor of a fluorescence microscope – blurring – can be captured as the Point Spread Function (PSF). Huygens uses this PSF to restore the original constituent object. As a result, image contrast and resolution are significantly improved, and noise decreased. Additional chromatic aberration, crosstalk, and unwanted movement can also be measured and corrected with Huygens. We will present Huygens as a complete package for the restoration, rendering, and analysis of images from a range of microscope types (widefield, confocal, spinning-disk, multi-photon, STED 3D). The latest advances in Huygens GPU-accelerated deconvolution and SPIM/Light Sheet image restoration will also be highlighted.
Presenter: Claire Weston, PhD
In this presentation we will describe a highly sensitive multiplex technique to visualize mRNA and miRNA in tissue sections and cultured cells. The beautiful images that result from this approach can be quantified using ImageDx, our proprietary image analysis software, to provide quantitative data at the single copy level. Learn how we can accelerate your research using a highly sensitive assay to visualize and quantify mRNA and miRNA in your biological samples.
Presenter Name and Title: David Bourdon, PhD. Senior Staff Scientist
Level: Intermediate level
To better understand human health and diseases, antibodies have been a critical tool for targeted detection of key biomarkers and proteins. By utilizing specific and sensitive antibodies as part of powerful Invitrogen immunoassay tools, we have enabled scientists to interrogate biological events such as cytokine and intracellular signaling cascades in a wide range of disease areas like cancer, autoimmunity, cardiovascular, and neurological diseases.
Invitrogen now offers a wide portfolio of antibody-based solutions including ELISA and multiplex assays on the Luminex platform. These tools enable accurate quantitation of specific proteins from sample types such as serum, plasma, and tissue culture supernatant. Come learn how these tools can augment your research, and may address common issues such as sample limitations and provide rich datasets for biomarker identification or pathway analysis.
Presenter: Mike Simson
Many of the antibodies published in biomedical research suffer from the lack of properly controlled studies into their reliability and reproducibility against specific antigens. Issues with improperly characterized antibodies prevented the replication of data in 47 of 53 preclinical studies. It is estimated that the resulting waste in labor and consumable costs are $350 million annually just for US researchers. In order to address this crisis two complimentary technologies have been developed that will create a more structured, accurate antibody system of validation. Bio-Layer Interferometry will be shown to determine Ab concentration, structural integrity, relative KD measurements, and epitope binning. Size exclusion chromatography multi-analyte platform will be shown to determine Ab off-site binding, specific binding, binding to protein complexes, and monomeric forms of proteins.
Presenter: Melanie Styers
Today’s scientific research is a global endeavor. Groundbreaking research is conducted in all parts of the world and by scientists of all nationalities. However, English remains the language of science, and the vast majority of scientific and medical journals are published in English. Thus, a lack of proficiency in English presents a significant barrier for acceptance of publications in these journals. EditBIOMED provides affordable scientific editing and proofreading services to help scientists publish their work in English-language journals. Our highly trained editors understand the complexities of the science, and also have an outstanding command of the English language. We correct the English, with an eye for the science, in order to help you publish your research.
Presenter: Andrew Moore, Editor-in-Chief
Many people click on but don’t fully read an article online; some don’t even find it. Many basically sound articles don’t even get to peer review. Which factors most influence the success of your manuscript, and at which stage, from submission to publication and final readership recognition? What are editors looking for? What are reviewers scrutinizing? Did you know that you’re writing for different readerships at different stages of your manuscript’s progress? How do search engines work, and how do editors work with authors to “optimize” their articles for search-engine-findability and general “noticeability”? Learn more about these topics, technological advances in publishing, and how you can contribute from the perspective of an author.
Presenter: Simon Fogarty – Director of Application Sciences
The Tecan D300e Digital Dispenser offers a simple method for rapidly generating compound serial dilutions, and compound combination or synergy experiments whilst minimising DMSO concentration in the assay. Using HP’s Direct Digital Dispensing technology, it provides picoliter to microliter non-contact dispensing of liquids directly into the cell plates, saving time, minimizing consumption of valuable samples and accelerating research. From small molecules in DMSO to biomolecules in surfactant-containing aqueous solutions, this convenient bench-top solution allows rapid delivery of any dose to any well. Requiring almost no set-up time, it uses disposable Dispense heads to minimize dead volumes and virtually eliminate the risk of cross-contamination, offering high quality, low volume dispensing for a wide range of applications.
Presenter: Viola Ruddat, Imaging Sales Specialist
Western blots have been around for over 30 years and are used in practically every life science laboratory. Equally well known, but often not satisfactorily addressed, are the challenges of reproducibility and the difficulty in obtaining accurate quantitative data and reliable results from Western blot experiments. This talk will describe the new Amersham WB system for SDS PAGE and Western blotting. Detection is based on fluorescence and the methods are standardized with built in evaluation software. The issues of reproducibility will be explored in more depth and how to obtain higher quality data will be discussed, with a focus on those key factors necessary to create consistent, quantifiable results.
Presenters: Paul Hung, Kurt Thorn
A major technical challenge for long-term analysis of cell behavior during in vitro culture is controlling and manipulating microenvironment parameters (temperature, gases, etc.) without moving the culture or impeding visualization.
In this Tech Talk workshop, microfluidic design engineer Dr. Paul Hung (EMD Millipore) and video microscopy center Director Prof. Kurt Thorn (UCSF) discuss in detail the latest advancements in microfluidically controlled cell culture and live cell microscopy. The speakers will discuss cell culture considerations, microfluidic design and fabrication requirements, and integrating microfluidic systems with microscopy.
The presenters then discuss a range of applications from bacterial biofilm dynamics to tumor metastasis to demonstrate cell shape analysis, individual cell tracking, and manipulation of the microenvironment followed by open discussion and a first look at “next generation” platforms.
Presenters: Carley Ross, Thomas Ramin
The extracellular vesicle (EV) research field has dramatically increased in the last five years. Using a high-speed flow cytometric sorter, EVs may be isolated at high rates such that researchers can differentially separate, isolate and characterize the EVs for downstream analysis. EVs contaminated with proteins, dye or antibody aggregates of the same size, but different mass, can be characterized based on these physical properties in the analytical ultra-centrifuge. We demonstrate that the XLA/I Analytical Ultracentrifuge (AUC) effectively separated particles on their sedimentation velocity and clarified issues with dye aggregation vs EV staining. Additionally, the DelsaMax Particle Characterizer allowed for quick analysis of post-sorted populations. The Astrios EQ Cell Sorter was able to sort EVs and AUC provided additional analysis for exosome purity.
Presenter: Colin Coates
We will discuss state-of-the-art, ultrasensitive, high speed camera technologies applications such as dynamic live cell imaging, single molecule detection and super-resolution microscopy.
sCMOS is a new high-performance imaging technology that can be considered progressive in its ability to simultaneously offer extremely low noise, rapid frame rates, wide dynamic range, high resolution and a large field of view, overcoming many performance trade-offs that are commonly associated with CCD detectors. EMCCDs however, retain the advantage of single photon sensitivity and often remain the best choice for extremely low light imaging.
This workshop will discuss and clarify the key sensor characteristics for consideration when selecting the optimum solution for your low light fluorescence microscopy application.
Presenter: Dr. Adam Elhofy, Ph.D., CSO, Essential Pharmaceuticals
Fetal bovine serum (FBS) has been used in cell culture for decades. Even though it introduces the greatest variability into cell culture, surprisingly FBS is not intensely scrutinized as a reagent. The use of FBS in cell culture applications will be examined, as its use may go against the basic principles of scientific experimentation in that the variables are not controlled for, it contains unknowns, and results from experiments using FBS in cell culture potentially cannot be replicated.
Presenter: Kevin Lowitz – Senior Product Manager, Protein & Cellular Analysis, Thermo Fisher Scientific
Western blotting remains one of the most ubiquitous techniques used by life science researchers around the world to advance their understanding of various disease areas such as cancer and immunology. Learn how new product innovations are providing more flexibility and better results than ever before to address a time-consuming and error-prone method for protein analysis.
We will discuss the new iBind™ Flex Western System to highlight:
- Sequential lateral flow technology for hands-free immunodetection
- Optimized platform for improved performance and potential cost savings
- Broad compatibility across the entire western workflow
Presenter: Daniel Appledorn
Cancer immunotherapy offers the potential for eradication of tumor cells and the prevention of cancer recurrence. In this study, we describe live cell imaging assays designed to quantify T cell killing of tumor cells in co-culture. PBMCs or isolated CD8+ T cells were added to tumor cells in combination with activators and a non-perturbing caspase-3/7 reagent in 96 or 384-well plates. Phase and 2-color images were captured using IncuCyte ZOOM™. Tumor cells and apoptotic cells were quantified using image analysis. These data demonstrate that a live cell imaging approach can discern the full time course and specificity of T cell killing, without lifting cells, using Ab labels or radioisotopes. HD images and time-lapse movies facilitate a clear understanding and compelling verification of the underlying biology.