Imaging Perspectives 2019

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- Cancer Research
  The field of cancer research spans diverse disciplines and requires a multipronged approach to understand the complex factors driving cancer formation and progression, as well as for the development of effective therapeutics.
  
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- Cell Counting
  The ability to accurately determine cell number is an important aspect of a broad range of applications, including setting up and optimizing cell-based assays, normalizing data across samples, and conducting cell proliferation assays.
  
  The Agilent BioTek Lionheart FX automated microscope and Agilent BioTek Cytation cell imaging multi-mode readers utilize label-free and fluorescence imaging modes, along with powerful Agilent BioTek Gen5 image analysis tools, to deliver efficient and effective automated cell counting.
- Cell Culture QC
  
  Automated imaging tools can provide valuable information for improving routine cell culturing techniques and increasing the effectiveness and reproducibility of downstream cell-based assays. Conventional methods for evaluating cell culturing techniques and assay design consist of manual inspection of a small subset of the cell population at random locations and time points.
- Cell Health and Viability
  
  Cell health and viability measurements provide essential insight into a broad range of biological processes and treatment responses. Diverse assay formats and reagents have been developed that measure specific aspects of cell viability corresponding to particular cellular response pathways and mechanisms of injury.
- Cell Metabolism
  
  Cellular metabolism comprises a number of biochemical reactions that occur in concert within the cells of living organisms. These orchestrated reactions, better described as metabolic pathways allow organisms to function and respond to environmental changes. Agilent cell metabolism assays detect discrete changes in cell bioenergetics in real time, providing a window into the critical functions that provide ATP, the energy that cells need for activation, signaling, proliferation, and biosynthesis.
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  Cell migration and invasion are live cell kinetic assays useful in oncology research. The principle difference between migration and invasion is that migration refers to normal cell movement; whereas invasion describes cells actively invading surrounding tissue. It can do this under the influence of a chemoattractant gradient and secretion of proteases. There are numerous methods for performing these assays including scratch assays performed on a monolayer of cells adhered to plasticware (microplates or culture inserts) or using 3D cell culture models.
- Developmental Biology
  A macro trend in drug discovery is the use of complex models of disease early in preclinical studies. An approach following this viewpoint is the use of whole organism models involving living invertebrates and vertebrates such as nematodes and zebrafish conducive to drug screening methods.
- Endotoxin Analysis
  Bacterial endotoxins are lipopolysaccharides (LPS) generally found in the outer membrane of gram-negative bacteria, such as E.coli, and are widely present in our environment. Because of the pyrogenic nature of endotoxins, with human effects from fever to hemorrhagic stroke, FDA regulations require that raw materials (e.g. water) used in parenteral drugs be tested for endotoxin levels.
- Histology and Slide Scanning
  Histology is typically performed on fixed tissue samples using upright microscopes equipped with oculars. Cytation and Lionheart FX provide automated digital imaging of these samples using color brightfield microscopy. Low to high magnifications from 1.25x to 60x (100x with Lionheart FX) can be combined with Cytation Gen5 image processing features such as image stitching to increase field of view.
- Immunology
  The immune system relies on diverse mechanisms working in concert to defend the host from infection and to identify and remove aberrant or damaged cells. Dysregulation of the immune system can lead to a variety of disease conditions, including autoimmune disorders, allergies, and cancer. Additionally, the immune system is a critical consideration for vaccine development and cell, tissue and organ transplants.
- Immuno-Oncology
  While harnessing the patient’s immune system to fight cancer is not a new concept, there have been significant strides in the past decade to more effectively direct and activate components of the immune system and counter the measures certain tumors adopt to shield itself from immune attack. These include immunotherapeutic drugs based on directing immune cells to tumors based on specific antigens they present, PD-1 checkpoint inhibition and the ex vivo manipulation of the patient’s own immune cells to better drive an immune response to cancer.
- Microbiology
  Since the advent of a lens system that enabled the visualization of bacteria and protists in 1671, the microscope remains a ubiquitous tool for the scientific study of live and fixed microorganisms. Investigating the structure, function, and typing of the highly diverse array of microorganisms spans heterogeneous applications including, but not limited to, the elucidation, diagnosis, and treatment of human and plant disease, food and water safety testing, vaccine development, investigation and production of industrial lipases, protein transfection to mammalian cells, environmental health and impact appraisal, horticultural and agricultural soil assessment, and exploration of alternative fuel sources.
- Molecular Biology and Biochemical
  Biochemical assays run the gamut from routine ELISAs to micro-volume nucleic acid and protein quantification. Endpoint and kinetic, single- and multi-wavelength assays are essential experimental tools. BioTek’s multi-mode readers have the flexibility to accommodate a very broad range of applications with superior performance.
- Rapid Cell Signaling
  G-protein coupled, seven-transmembrane (7-TM) receptors (GPCRs) represent large, diverse gene families found in fungi, plants and animals. The GPCR family of receptors is involved primarily with internalizing extracellular environmental signals via interaction with intracellular G-proteins. GPCR ligands and modulators range from peptide and non-peptide molecules to chemokines, nucleosides, growth factors and light.
- Stem Cell Research
  Research into the reprogramming of somatic cells into pluripotent stem cells received the Nobel Prize in 2012. The long term importance of this research resides in the use of stem cells for regenerative therapies; for the short term, however these cells represent a potential unlimited source of highly physiologically relevant cells, akin to human primary cells, that can be used as drug discovery reagents.
- Virology
  Research into the diverse aspects of virology are made possible by a broad range of applications and assay formats, from relatively simple methods for viral detection to those designed to elucidate complex immune responses and pathological processes. BioTek instruments provide versatile platforms for conducting viral research, with optimized imaging- and plate reader-based solutions, as well as liquid handling and automation tools for increased assay throughput and reproducibility.
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Elison Blancaflor
Noble Research Institute LLC

Description: Cross section of a pecan root stained with calcofluor to label the cellulosic cell wall (blue color). White/pink color is autofluorescence that indicates vascular cells with enhanced wall thickening. The image was captured at 10x and with brightfield and DAPI, FITC and Rhodamine filters.
Image captured with: Lionheart FX

Ibrahim Halil Demirsoy
Luigi Vanvitelli Campania University

Description: Stem cell -derived neural cells imaged at 40x magnification, Actin (green), MitoTracker (red) and DAPI (blue).
Image captured with: Cytation 3

Rene Girard
University of Texas Health Science Center at Tyler

Description: Rabbit lung, 20x, Manual mode with DAPI, GFP, and 647 Filters, Image shows rabbit lung injury caused by acute empyema. Staining shows Nuclear (DAPI), Fibrin (Red), and PAI-1 (Green) co-localization on the lining of the lung. Empyema causes fibrin formation in the pleural space.
Image captured with: Cytation 5

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