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What is Live-Cell Imaging?

Besides the structural organization of cells or organs, dynamic processes are a major contributor to a functioning biological entity. Naturally, these processes can be best observed in living cells with non-invasive techniques like optical methods, collectively called “live-cell imaging” methods. Live-cell imaging covers all techniques where live cells are observed with microscopes – from the observation of embryogenesis with stereo microscopes, via cell growth studies with compound microscopes, until studies of physiological states of cells or cellular transport using fluorescent dyes or proteins. Although being highly demanding for both, experimenter and equipment (e.g. imaging systems, climate control), live-cell imaging techniques deliver results that are indispensable for present-day research.

  • Deconvolution

    Fluorescence microscopy is a modern and steadily evolving tool to bring light to current cell biological questions. With the help of fluorescent proteins or dyes it is possible to make discrete cellular components visible in a highly specific manner. A prerequisite for these kinds of investigations is a powerful fluorescence microscope. One special aim is the three-dimensional illustration of a structure to get an impression of full plasticity. This poses a certain problem for the experimenter using a classical light microscope.
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  • Webinar: Live Cell Imaging

    Cells communicate and interact with one another to transmit signals and initiate reactions that facilitate coordinated events. These interactions are critical in almost every aspect of physiology, from transmitting neuronal signals that allow us to sense hot and cold to initiating immune responses that fight against infection.
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  • Postlipolytic Insulin-dependent Remodeling of Micro Lipid Droplets in Adipocytes

    Despite the lipolysis–lipogenesis cycle being a fundamental process in adipocyte biology, very little is known about the morphological changes that occur during this process. The remodeling of lipid droplets to form micro lipid droplets (mLDs) is a striking feature of lipolysis in adipocytes, but once lipolysis ceases, the cell must regain its basal morphology.
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  • Scientific Illustration and Animation for Research Scientists

    One of the crucial tasks of a research scientist is reporting and communicating about his work. This is vital for cutting edge research; it is crucial to gain insights from other experts, to get a discussion going, to improve, to be able to get some funding, to convince other colleagues that your research is good. Imagine that communication is not there. Then you will immediately appreciate its importance. There would be a lot of individuals all having a single piece of the puzzle in their hands, but nobody seeing the complete picture.
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  • Live-Cell Imaging

    The understanding of complex and fast cellular dynamics is an important step to get insight into biological processes. Therefore, today’s life science research more and more demands studying physiological events on the molecular level in real-time.
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  • Protein Transport Processes at the Apical Membrane of Polarized Epithelial Cells

    Due to their special role in organ function and the exchange of biological components some body cells developed certain polarization characteristics. These are reflected in differences of their plasma membrane composition. The essential and fascinating task of polarized protein transport in epithelial cells is to get the right protein into the right membrane.
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  • Total Internal Reflection Fluorescence (TIRF) Microscopy

    Total internal reflection fluorescence (TIRF) is a special technique in fluorescence microscopy developed by Daniel Axelrod at the University of Michigan, Ann Arbor in the early 1980s. TIRF microscopy delivers images with an outstandingly high axial resolution below 100 nm. This allows the observation of membrane-associated processes.
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  • Applications of TIRF Microscopy in Life Science Research

    The special feature of TIRF microscopy is the employment of an evanescent field for fluorophore excitation. Unlike standard widefield fluorescence illumination procedures with arc lamps, LEDs or lasers, the evanescent field only penetrates the specimen by about 100 nm starting from the coverslip/medium interface.
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  • Webinar: Morphogenesis

    Morphogenesis – literally “shape creation” – is responsible for the diversity of biological shapes that make up Darwin’s “endless forms most beautiful and wonderful”. In recent years, the combination of cutting edge microscopy and molecular approaches in developmental, cell, and molecular biology have provided an increasingly in-depth view of how organisms (and all of their integral parts) form from a single cell. In this exciting webinar...
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  • Live-cell Imaging Techniques

    The understanding of complex and/or fast cellular dynamics is an important step for exploring biological processes. Therefore, today’s life science research is increasingly focusing on dynamic processes like cell migration, morphological changes of cells, organs or whole animals and physiological (e.g. changes of intracellular ion composition) events in living specimens in real time.
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  • The New Repository on the Block

    The need for data validation and accessibility has never been greater than it is today. We are inundated with information from a multitude of resources, but how can we easily evaluate the accuracy of that data? In the past, the peer review process provided this and was often run by publishers.
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  • Differential Interference Contrast

    The examination of live unstained biological specimens often suffers from poor contrast and therefore bad visibility of the specimen. Thick specimens in particular, such as brain slices, show up as nothing more than light grey structures instead of single cells. This tutorial explains the optical elements in the light path and the operating mode of DIC (differential interference contrast) on the example of an inverted and motorized high-end research light microscope which can be used for transmitted light contrasting methods and fluorescence microscopy.
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  • Ratiometric Imaging

    Many fundamental functions of a cell strongly depend on delicate, but nevertheless dynamic balances of ions (e.g. calcium, magnesium), voltage potentials and pH between the cell’s cytosol and the surrounding extracellular space. Ratiometric imaging allows reliable estimations of ion concentrations and pH or voltage changes by measuring fluorophore emission shifts.
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  • Ratiometric Imaging Needs a Specialized Setup

    Ratiometric imaging is widely used to study highly dynamic intracellular ion, voltage or pH changes. The most common application, however, is calcium imaging. Ratiometric imaging is also used for investigating cellular networks, where e.g. relative calcium concentrations are passed among cells or different cell types dynamically change ion, voltage or pH levels. Also FRET assays can considerably benefit from ratiometric imaging, as the signal-to-noise-ratio is greatly improved.
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  • The Patch-Clamp Technique

    Especially in neuroscience, the physiology of ion channels has always been a major topic of interest. The development of the patch-clamp technique in the late 1970s has given electrophysiologists new prospects. It allows high-resolution current recordings not only of whole cells, but also of excised cellular patches. Even single-channel opening events can be investigated. However, with its complex technical, physical and biological background, the need for highly sensitive equipment and the huge amount of skills required of the experimenter, electrophysiology is still one of the most challenging methods in daily laboratory work.
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  • Step by Step Guide to Hybrid Detection and Photon Counting

    This tutorial explains the underlying hybrid detection technology and compares it to photomultiplier technology. The implications of hybrid detection design for imaging and photon counting are discussed. The tutorial closes with a brief summary of photon counting in the context of imaging.
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  • STED Nanoscopy of Actin Dynamics in Synapses deep inside Living Brain Slices

    It is difficult to investigate the mechanisms that mediate long-term changes in synapse function because synapses are small and deeply embedded inside brain tissue. Although recent fluorescence nanoscopy techniques afford improved resolution, they have so far been restricted to dissociated cells or tissue surfaces. However, to study synapses under realistic conditions, one must image several cell layers deep inside more-intact, three-dimensional preparations that exhibit strong light scattering, such as brain slices or brains in vivo.
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  • Nanoscopy in a Living Multicellular Organism Expressing GFP

    We report superresolution fluorescence microscopy in an intact living organism, namely Caenorhabditis elegans nematodes expressing green fluorescent protein (GFP)-fusion proteins. We also superresolve, by stimulated emission depletion (STED) microscopy, living cultured cells, demonstrating that STED microscopy with GFP can be widely applied.
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  • Webinar: Principles and Applications of 
Live Cell Imaging

    Live cell imaging has become a cornerstone of modern cell biology, as well as a technique routinely employed in other fields such as developmental biology, stem cell biology and neuroscience. In this webinar, you will learn about the effective use of live cell imaging for a growing number of applications.
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  • TIRF Microscopy of the Apical Membrane of Polarized Epithelial Cells

    Application of TIRF microscopy (Total Internal Reflection Fluorescence) allows the visualization of structures at the apical surface of polarized epithelial cells that have been hidden in conventional fluorescence microscopy images. Hence, the approach reveals new insights into the composition of this characteristic cell pole that elucidate processes in apical protein trafficking.
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  • Mapping Billions of Synapses with Microscopy and Mathematics

    A combination of widefield imaging techniques and image segmentation analysis enable researchers to map learning-induced functional changes in individual synapses throughout the hippocampus.
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  • Webinar: Forces in Cell Biology

    Chemical and electrical signals are well known to contribute to the growth and development of an organism; recently, there has been an emerging focus on another cue that can inform cellular development: physical force. Cells are pushed, pulled, and squeezed as they undergo biological processes such as cell division, migration, and morphogenetic events, and exciting recent work is identifying the role such forces play in governing cell function.
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  • Sniffing Out the Secrets of Social Behavior

    Yet we are only just beginning to understand the complexities and functional differences of the sense of smell in mammals. Prof. Marc Spehr, head of the Department of Chemosensation at RWTH Aachen University since 2009, explains his findings on the neuronal mechanisms of olfactory perception and signal processing using the mouse model. He and his team are trying to find out how substances for social interaction are perceived and how this perception generates a specific type of behavior.
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  • Exploring the Concert of Neuronal Activities

    Brain research using Confocal and Multiphoton Microscopy. Using imaging techniques such as confocal and two-photon microscopy, neuronal dendritic arborization of neurons and their synaptic interconnections can be visualized.
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  • Stem Cell Biology in Cancer Research

    The comprehension of stem cell biology and its molecular basis is now acquiring paramount importance in cancer research. The need to look at a single, possibly living, cell makes fluorescence microscopy and confocal microscopy invaluable allies in the study of stem cells.
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  • New Standard in Electrophysiology and Deep Tissue Imaging

    The function of nerve and muscle cells relies on ionic currents flowing through ion channels. These ion channels play a major role in cell physiology. One way to investigate ion channels is to use patch clamping. This method allows investigation of ion channels in detail and recording of the electric activity of different types of cells, mainly excitable cells like neurons, muscle fibres or beta cells of the pancreas. The patch clamping technique was developed by Erwin Neher and Bert Sakmann in the 1970s and 80s to study individual ion channels in living cells. In 1991 they received the Nobel Prize for Physiology and Medicine for their work. Today the patch clamping technique is one of the most important methods in the field of electrophysiology.
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  • Exploring Cell Logistics

    Using TIRF microscopy, scientists have been able to take a closer look at intracellular transport processes with the example of the galactose-binding protein Galectin-3, which has been identified as a potential apical sorting receptor.
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Directory of open access journals
The EMBO Journal
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Journal of Cell Science
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Journal of Biophotonics
International, peer-reviewed, open-access, online publication
Proceedings B - the Royal Society's biological research journal
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