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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
The First Supercontinuum Confocal that Adapts to the Sample
Until now, biological and medical research fluorescence imaging in multi-user facilities or institutes has been limited by the type or number of dyes that could be excited. The Leica TCS SP5 X…Read article
Deep Tissue Imaging
Developmental biology using Multiphoton microscopy with OPO. To gain new insight into the fundamental control of cell response to physical changes and to study the dynamics and roles of biological…Read article
ATP Changes the Fluorescence Lifetime of Cyan Fluorescent Protein via an Interaction with His148
This study employed fluorescence lifetime spectroscopy to investigate the influence of ATP (adenosine triphosphate) on the fluorescence intensities of yellow and cyan fluorescent proteins (YFP/CFP)…Read article
Choose Your Excitation Wavelength
Although time correlated single photon counting (TCSPC) is the method of choice for fluorescence lifetime quantification, it requires dedicated instrumentation including a pulsed laser source, a…Read article
A mTurquoise-Based cAMP Sensor for Both FLIM and Ratiometric Read-Out Has Improved Dynamic Range
FRET-based sensors for cyclic Adenosine Mono Phosphate (cAMP) have revolutionized the way in which this important intracellular messenger is studied. The currently prevailing sensors consist of the…Read article
An Introduction to Fluorescence
Fluorescence is widely used in microscopy and an important tool for observing the distribution of specific molecules. Most molecules in cells do not fluoresce. They therefore have to be marked with…Read article
A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms
Electron microscopy (EM) achieves the highest spatial resolution in protein localization, but specific protein EM labeling has lacked generally applicable genetically encoded tags for in situ…Read article
Modern Fluorescent Proteins and their Biological Applications
Here we present two review articles on fluorescent proteins and their biological applications. These first article reviews our current knowledge of blue, green, and red chromophore formation in…Read article
Fluorescent Proteins – Introduction and Photo Spectral Characteristics
The prospects of fluorescence microscopy changed dramatically with the discovery of fluorescent proteins in the 1950s. The starting point was the detection of the jellyfish Aequorea victoria green…Read article
Handbook of Optical Filters for Fluorescence Microscopy
Fluorescence microscopy and other light-based applications require optical filters that have demanding spectral and physical characteristics. Often, these characteristics are application-specific and…Read article
Sample Preparation for GSDIM Localization Microscopy – Protocols and Tips
The widefield super-resolution technique GSDIM (Ground State Depletion followed by individual molecule return) is a localization microscopy technique that is capable of resolving details as small as…Read article
Imaging Pheromone Sensing in a Mouse Vomeronasal Acute Tissue Slice Preparation
In mice, the ability to detect pheromones is principally mediated by the vomeronasal organ (VNO). Here, an acute tissue slice preparation of VNO for performing calcium imaging is described. This…Read article
Colonization of Potato Rhizosphere by GFP-Tagged Bacillus subtilis MB73/2, Pseudomonas sp. P482 and Ochrobactrum sp. A44 Shown on Large Sections of Roots Using Enrichment Sample Preparation and Confocal Laser Scanning Microscopy
The ability to colonize the host plants’ rhizospheres is a crucial feature to studyin the case of Plant Growth Promoting Rhizobacteria (PGPRs) with potential agricultural applications. In this work,…Read article
Spectral and Structural Comparison Between Bright and Dim Green Fluorescent Proteins in Amphioxus
The cephalochordate Amphioxus naturally co-expresses fluorescent proteins (FPs) with different brightness, which thus offers the rare opportunity to identify FP molecular feature/s that are associated…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
Correlative In-Resin Super-Resolution and Electron Microscopy Using Standard Fluorescent Proteins
We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer…Read article
Fluorescent Proteins Illuminate Cell Biology
Green fluorescent protein (GFP) isolated from the jellyfish Aequorea victoria and GFP-like fluorescent proteins from other animals have had an important role in the technical innovations that have…Read article
Video Talk by Roger Tsien: Fluorescent Proteins
Live cell imaging has been revolutionized by the discovery of the green fluorescent protein (GFP). This lecture covers the history of GFP, how it folds and becomes fluorescent, how it has been mutated…Read article
Video Talk by Roger Tsien: Fluorescent Protein Indicators
In this talk, Roger Tsien discusses how fluorescent proteins have been turned into indicators for a wide variety of biological molecules, including pH, ions, redox potential, and signaling molecules…Read article
Clap On, Clap Off. Protein On, Protein Off
Unlike genetic or pharmacological manipulation, light-controlled proteins respond immediately, can be temporally and spatially triggered, are reversible, and are specific to the protein of interest.…Read article
Photoactivatable, photoconvertible, and photoswitchable Fluorescent Proteins
Fluorescent proteins (FPs) such as GFP, YFP or DsRed are powerful tools to visualize cellular components in living cells. Nevertheless, there are circumstances when classical FPs reach their limits.…Read article
What is Photomanipulation?
The term photomanipulation describes a wide range of techniques that enable the microscopist the transition from passive observer to instigator of events by offering a way of interacting with their…Read article
Real Time Observation of Neutrophil White Blood Cell Recruitment to Bacterial Infection In Vivo
The zebrafish (Danio rerio) is an emerging vertebrate model organism to study infection. The transparent larva comprises a fully functional innate immune system and enables live imaging of fluorescent…Read article
The Fundamentals and History of Fluorescence and Quantum Dots
At some point in your research and science career, you will no doubt come across fluorescence microscopy. This ubiquitous technique has transformed the way in which microscopists can image, tag and…Read article
FLIM FRET and Biosensors: Versatile Tools for Biomedical Research
Fluorescence Lifetime Imaging (FLIM) in combination with Förster Resonance Energy Transfer (FRET) has proven to be very beneficial for investigations in biomedical research for a wide range of…Read article