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4D Photoactivation of pa-GFP in Living Cells Using Two-Photon Excitation Laser Scanning Microscopy
We report about two-photon activation of a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (pa-GFP). This special form of the molecule increases its fluorescence…Read article
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…Read article
In Vivo Labeling Method Using a Genetic Construct for Nanoscale Resolution Microscopy
We demonstrate beam scanning-stimulated emission depletion microscopy with in vivo labeled cells. A red emitting fluorescent dye is introduced into membrane protein fused to a multifunctional reporter…Read article
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…Read article
FRET Sensitized Emission Wizard Widefield
Förster Resonance Energy Transfer (FRET) is a technique, which allows insight into the interactions between proteins or molecules in proximities beyond light microscopic resolution. An excited…Read article
Selective Plane Illumination Microscopy Techniques in Developmental Biology
Selective plane illumination microscopy (SPIM) and other fluorescence microscopy techniques in which a focused sheet of light serves to illuminate the sample have become increasingly popular in…Read article
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…Read article
Light Sheet Fluorescence Microscopy - A Review
Light sheet fluorescence microscopy (LSFM) functions as a non-destructive microtome and microscope that uses a plane of light to optically section and view tissues with subcellular resolution. This…Read article
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…Read article
Optical Contrast Methods
Optical contrast methods give the potential to easily examine living and colorless specimens. Different microscopic techniques aim to change phase shifts caused by the interaction of light with the…Read article
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…Read article
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…Read article
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…Read article
Quantitative Imaging in Cell Biology: Light Sheet Microscopy
This chapter introduces the concept of light sheet microscopy along with practical advice on how to design and build such an instrument. Selective plane illumination microscopy is presented as an…Read article
Two-color STED Microscopy of Living Synapses using a Single Laser-beam Pair
The advent of superresolution microscopy has opened up new research opportunities into dynamic processes at the nanoscale inside living biological specimens. This is particularly true for synapses,…Read article
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…Read article
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…Read article
Fluorescence 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 focused on dynamic…Read article
Introduction to 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…Read article
Controlling the TIRF Penetration Depth is Mandatory for Reproducible Results
The main feature of total internal reflection fluorescence (TIRF) microscopy is the employment of an evanescent wave for the excitation of fluorophores instead of using direct light. A property of the…Read article
Digital Camera Technologies for Scientific Bio-Imaging
This four-part series of articles published in Microscopy and Analysis covers the factors to consider in choosing a camera among CCD, EMCCD, and scientific-grade CMOS camera technologies for…Read article
Light Sheet Fluorescence Microscopy: Beyond the Flatlands
Light Sheet Fluorescence Microscopy (LISH-M) is a true fluorescence optical sectioning technique, first described by Heinrich Siedentopf in 1902 under the name of Ultramicroscopy. Light sheet…Read article
Imaging Enzymes at Work
For the understanding of functions of proteins in biological and pathological processes, reporter molecules such as fluorescent proteins have become indispensable tools for visualizing the location of…Read article
Direct In Vivo Evidence for Tumor Propagation by Glioblastoma Cancer Stem Cells
Cancer research using multiphoton microscopy. High-grade gliomas (World Health Organization grade III anaplastic astrocytoma and grade IV glioblastoma multiforme), the most prevalent primary…Read article
A Precise and Rapid Mapping Protocol for Correlative Light and Electron Microscopy of small invertebrate organisms
CLEM (correlative live cell and electronmicroscopy) seeks to bridge the data acquired with different imaging strategies, typically between light microscopy and electron microscopy. It has been…Read article
New Labeling Tools Can Help to Realize the Full Potential of Super-Resolution Microscopy
Since super-resolution microscopy techniques revolutionized the concept of light microscopy by overcoming the physical diffraction limit, STED microscopy and other super-resolution techniques have…Read article
Watching Molecule Movements in Live Cells
The newly developed STED-RICS microscopy method records rapid movements of molecules in live samples. By combining raster image correlation spectroscopy (RICS) with STED fluorescence microscopy,…Read article
From Dynamic Live Cell Imaging to 3D Ultrastructure: Novel Integrated Methods for High Pressure Freezing and Correlative Light-Electron Microscopy
To correlate dynamic events in adherent cells with both ultrastructural and 3D information, we developed a method for cultured cells that combines confocal time-lapse images of GFP-tagged proteins…Read article
A Flip of the Mitotic Spindle has Disastrous Consequences for Epithelial Cells
Constructing a body is like building a house – if you compromise structural integrity, the edifice can collapse. Nowhere is that clearer on a cellular level than in the case of epithelial sheets,…Read article
Nobel Prize 2013 in Physiology or Medicine for Discoveries of the Machinery Regulating Vesicle Traffic
On October 7th 2013, The Nobel Assembly at Karolinska Institutet has decided to award The Nobel Prize in Physiology or Medicine 2012 jointly to James E. Rothman, Randy W. Schekman and Thomas C. Südhof…Read article
Live-Cell Imaging Evolves to Find New Niches
Since its introduction in the 1600s, improvements in microscope technology have continually broadened the types of cells and cellular processes that can be studied. Advances in automation have made…Read article
Laser Microdissection (LMD) and Fancy Applications
New and far-reaching applications have recently been opened up in the field of laser microdissection. In addition to conventional dissection, the laser microdissection system (LMD) of Leica…Read article
The Environment Makes the Stem Cell
A recent publication in Nature shows that all stem cells divide and compete for niche space by passively "kicking out" others so that eventually one stem cell takes over the whole niche. Jacco van…Read article
STED Microscopy of Living Cells – New Frontiers in Membrane and Neurobiology
Recent developments in fluorescence far-field microscopy such as STED microscopy have accomplished observation of the living cell with a spatial resolution far below the diffraction limit. Here, we…Read article
Nanoscale Protein Diffusion by STED-Based Pair Correlation Analysis
We describe for the first time the combination between cross-pair correlation function analysis (pair correlation analysis or pCF) and stimulated emission depletion (STED) to obtain diffusion maps at…Read article
Axon Injury and Regeneration in the Adult Drosophila
Neural regeneration is a fascinating process with profound impact on human health, such that defining biological and genetic pathways is of interest. Here we describe an in vivo preparation for…Read article
"Leica is always flexible and dynamic" - Interview with Audrey Salles, Pasteur Institute, Paris
Audrey Salles is a specialist for confocal and super-resolution microscopy at Pasteur Institute, Imagopole, PFID, Paris, France. Her research interests are cytokine signaling and skeleton organization…Read article
Nature Methods: Light-sheet Fluorescence Microscopy - Method of the Year 2014
Just about everyone who has examined fluorescent samples under the microscope is aware of the constant struggle to have enough signal to see the labeled structures while also avoiding fluorophore…Read article
Video: High Speed Scanning – With two Scanners in one System
High speed scanning is necessary to image rapidly changing biological processes. With traditional scanning techniques, imaging speed is limited by the number of fluorophores in a specimen. And, rapid…Read article
Universal PAINT – Dynamic Super-Resolution Microscopy
Super-resolution microscopy techniques have revolutionized biology for the last ten years. With their help cellular components can now be visualized at the size of a protein. Nevertheless, imaging…Read article
Third Harmonic Generation Microscopy – Label-Free 3D-Tissue Imaging and Blood Flow Characterization
THG microscopy as special variants of multiphoton microscopy. Third Harmonic Generation (THG) microscopy is a non-fluorescent multi-photon technique that combines the advantages of label-free…Read article
Correlative Light and Electron Microscopy – Get Your Free CLEM e-Book for Download
The urge to go deeper into the microscopic world has led researchers to combine the versatility of the light microscopy (LM) with the resolution power of the electron microscope (EM) to produce…Read article
Multi-protein Assemblies Underlie the Mesoscale Organization of the Plasma Membrane
Most proteins have uneven distributions in the plasma membrane. Broadly speaking, this may be caused by mechanisms specific to each protein, or may be a consequence of a general pattern that affects…Read article
Cortical Actin Networks Induce Spatio-temporal Confinement of Phospholipids in the Plasma Membrane – A Minimally Invasive Investigation by STED-FCS
Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of…Read article
Probes that FIT RNA
We have been developing new tools based on fluorogenic forced intercalation (FIT) probes for RNA detection quantification and interference in biological samples. Upon duplex formation with target…Read article
Light Sheet Microscopy Turned Vertically
Living cells and organisms often suffer from the high light intensities used for fluorescent imaging. Light sheet microscopy reduces phototoxic effects and bleaching by illuminating a specimen in only…Read article
FRAP with TCS SP8 Resonant Scanner
Fast FRAP experiments need a sufficient number of measurement points for meaningful interpretation and fitting analysis. To study very fast translocational processes, the use of a resonant scanner…Read article
4Pi-RESOLFT Nanoscopy
Here we apply the 4Pi scheme to RESOLFT nanoscopy using two-photon absorption for the on-switching of fluorescent proteins. We show that in this combination, the lobes are so low that low-light level,…Read articleHow to do a Proper Cell Culture Quick Check
In order to successfully work with mammalian cell lines, they must be grown under controlled conditions and require their own specific growth medium. In addition, to guarantee consistency their growth…Read article
What Makes sCMOS Microscope Cameras so Popular?
sCMOS cameras are more sensitive and are capable of much higher acquisition speed than cameras with other sensor types. Even though CCD cameras are widely used in live cell imaging and time-lapse…Read article
The Bimodally Expressed MicroRNA miR‐142 Gates Exit from Pluripotency
A stem cell's decision to self‐renew or differentiate is thought to critically depend on signaling cues provided by its environment. It is unclear whether stem cells have the intrinsic capacity to…Read article
Individual Macromolecule Motion in a Crowded Living Cell
There is solid evidence for analyzing fluorescence correlation and dual color fluorescence crosscorrelation spectroscopy data ( FCS and dual color FCCS) in cellular applications by equations based on…Read article
Label-free in vivo Imaging of Myelinated Axons in Health and Disease with Spectral Confocal Reflectance Microscopy
We report a new technique for high-resolution in vivo imaging of myelinated axons in the brain, spinal cord and peripheral nerve that requires no fluorescent labeling. This method, based on spectral…Read article
Highly Selective Fluorescent and Colorimetric Probe for Live-cell Monitoring of Sulphide Based on Bioorthogonal Reaction
H2S is the third endogenously generated gaseous signaling compound and has also been known to involve a variety of physiological processes. To better understand its physiological and pathological…Read article
Adeno-associated Viral Vectors do not Efficiently Target Muscle Satellite Cells
Adeno-associated viral (AAV) vectors are becoming an important tool for gene therapy of numerous genetic and other disorders. Several recombinant AAV vectors (rAAV) have the ability to transduce…Read article
Super-Resolution Optical Microscopy of Lipid Plasma Membrane Dynamics
Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence…Read article
Chronic Inflammation Under the Microscope
In the course of chronic inflammation certain body areas are recurrently inflamed. This goes along with many human diseases. With the help of widefield light microscopy, the underlying processes can…Read article
Axial Tubule Junctions Control Rapid Calcium Signaling in Atria
The canonical atrial myocyte (AM) is characterized by sparse transverse tubule (TT) invaginations and slow intracellular Ca2+ propagation but exhibits rapid contractile activation that is susceptible…Read article
Introduction to Mammalian Cell Culture
Mammalian cell culture is one of the basic pillars of life sciences. Without the ability to grow cells in the lab, the fast progress in disciplines like cell biology, immunology, or cancer research…Read article
Acute Transcriptional Up-regulation Specific to Osteoblasts/Osteoclasts in Medaka Fish Immediately after Exposure to Microgravity
Bone loss is a serious problem in spaceflight; however, the initial action of microgravity has not been identified. To examine this action, we performed live-imaging of animals during a space mission…Read article
Solvent immersion imprint lithography
We expand upon our recent, fundamental report on solvent immersion imprint lithography (SIIL) and describe a semi-automated and high-performance procedure for prototyping polymer microfluidics and…Read article
Live Cell Isolation by Laser Microdissection
Laser microdissection is a tool for the isolation of homogenous cell populations from their native niches in tissues to downstream molecular assays. Beside its routine use for fixed tissue sections,…Read article
Kaggle Competition for Multi-label Classification of Cell Organelles in Proteome Scale Human Protein Atlas Data
The Cell Atlas, a part of the Human Protein Atlas, was created by the group of Prof. Emma Lundberg at the SciLifeLab, KTH Royal Institute of Technology, in Stockholm, Sweden. Currently, she is a…Read article
Extending Nanoscopy Possibilities with STED and exchangeable fluorophores
When it comes to STED Nanoscopy, keeping high signal-to-noise is key to achieve the best possible resolution in fixed and living cells. This can be challenging in the case of experiments in 3D and/or…Read article
Development of fluorescence lifetime imaging microscopy (FLIM) and its relevance for functional imaging
Prof. Ammasi Periasamy, Director, Keck Center for Cellular Imaging, University of Virginia, was interviewed by Dr. Giulia Ossato, Product Manager functional imaging, during Leica Microsystems Meets…Read article
Imaging Intracellular Temperature using Fluorescence Lifetime Imaging Microscopy (FLIM)
This video shows the presentation of Dr. Kohki Okabe given at the 9th Congress of the Federation of Asian and Oceanian Physiological Society (FAOPS) held in Kobe, Japan on 30 March 2019. Dr. Okabe…Read article
New Light Shed on the Nanodomain Organization of the Endoplasmic Reticulum (ER)
The endoplasmic reticulum (ER) is a continuous membrane organelle in charge of protein synthesis, lipid synthesis and detoxification. The ER structure is described in terms of smooth peripheral…Read article
Step by Step Guide to the Molecular Basics of CARS Microscopy
CARS (Coherent Anti-Stokes Raman Scattering) microscopy is a dye-free method which images structures by displaying the characteristic intrinsic vibrational contrast of their molecules. The crucial…Read article
How to prepare samples for Stimulated Raman Scattering
Stimulated Raman Scattering (SRS) is an emerging microscopy technique that images the vibrations of chemical bonds within a specimen. It allows researchers to probe the endogenous chemistry or…Read article
Improvement of Imaging Techniques to Understand Organelle Membrane Cell Dynamics
Understanding cell functions in normal and tumorous tissue is a key factor in advancing research of potential treatment strategies and understanding why some treatments might fail. Single-cell…Read article
Following Ca²⁺ Changes in Acutely Stimulated Spheroids
Spheroids are easy and cost-effective 3D cell cultures, useful for fundamental and applied research. They can bridge the gap between 2D and in vivo experiments and can be used for high-throughput…Read article
Coherent Raman Scattering Microscopy Publication List
CRS (Coherent Raman Scattering) microscopy is an umbrella term for label-free methods that image biological structures by exploiting the characteristic, intrinsic vibrational contrast of their…Read article