Learn & Share: Widefield Microscopy?
Widefield microscopy refers to a basic sample illumination principle in microscopy. Since widefield microscopy permanently illuminates the whole sample, it can be distinguished from confocal microscopy where only one single focal spot is illuminated and recorded at a time. Typically, widefield microscopy utilizes light sources such as halogen, metal halide lamps or LED for sample illumination. Detection is performed through oculars or with the help of a digital camera. Several contrast methods increase widefield microscopy capabilities, starting from Phase Contrast through to Differential Interference Contrast (DIC) or fluorescence, to name but a few. Computer based deconvolution can be applied to increase widefield fluorescence image quality and enable 3D image reconstruction. Moreover, TIRF and GSDIM super-resolution microscopy can be assigned to widefield microscopy.
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Image Gallery: THUNDER Imager
To help you answer important scientific questions, THUNDER Imagers eliminate the out-of-focus blur that clouds the view of thick samples when using camera-based fluorescence microscopes. They achieve…Read article -
How to Develop an Airway Epithelial Cell Model
The present study describes the optimal growth conditions for the proliferation and differentiation of bovine bronchial epithelial cells (BBECs). The cells are grown at an air-liquid interface (ALI)…Read article -
THUNDER Technology Note
So far, widefield microscopy was not suitable to image larger volumes, since the contrast of the recorded fluorescence image is reduced by the background (BG) mainly originating from out of focus…Read article -
Proline Metabolism is Essential for Trypanosoma brucei brucei Survival in the Tsetse Vector
Bloodsucking insects play a major role in the transmission of pathogens that cause major tropical diseases. Their capacity to transmit these diseases is directly associated with the availability and…Read article -
mTORC1 Promotes Proliferation of Immature Schwann Cells and Myelin Growth of Differentiated Schwann Cells
The myelination of axons is essential for neuronal wiring and normal nervous system functions. In the peripheral nervous system, Schwann cells (SCs) form myelin sheaths around axons during nerve…Read article -
Effect of curcumin analogs onα-synuclein aggregation and cytotoxicity
Alpha-synuclein (α-Syn) aggregation into oligomers and fibrils is associated with dopaminergic neuron loss occurring in Parkinson’s disease (PD) pathogenesis. Compounds that modulate α-Syn aggregation…Read article -
Expression Analysis of Platelet‐derived Growth Factor Receptor Alpha and its Ligands in the Developing Mouse Lung
Activation of the platelet‐derived growth factor receptor‐α (PDGFR α) signaling pathway is critically important during lung alveogenesis, the process in lung development during which alveoli are…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 -
Adipose Stem Cells Display Higher Regenerative Capacities and More Adaptable Electro-Kinetic Properties Compared to Bone Marrow-Derived Mesenchymal Stromal Cells
Adipose stem cells (ASCs) have recently emerged as a more viable source for clinical applications, compared to bone-marrow mesenchymal stromal cells (BM-MSCs) because of their abundance and easy…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 -
Introduction to Widefield Microscopy
One of the most basic microscopy techniques is known as ‘Widefield Microscopy’. It is fundamentally any technique in which the entire specimen of interest is exposed to the light source with the…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 -
Image Gallery: Application Options with Leica DM2500 LED Microscope
The number of applications in microscopy can be as manifold as life itself. Different specimens demand different contrast methods to get a decent result. Unstained cells normally do not show a very…Read article -
Definitions of Basic Technical Terms for Digital Microscope Cameras and Image Analysis
Most microscopes today are operated with a camera. The characteristics of the camera often decide whether the acquired image will reveal what a researcher wants to see. But when diving into camera…Read article -
How 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 -
Infinity Optical Systems
“Infinity Optics” refers to the concept of a beam path with parallel rays between the objective and the tube lens of a microscope. Flat optical components can be brought into this “Infinity Space”…Read article -
Introduction to Digital Camera Technology
A significant majority of modern optical microscopy techniques require the use of a digital camera. By working with digital devices researchers can observe specimens on a screen in real time or…Read article -
Color Infidelity: Why Using a Light Source Incorrectly is Cheating on your Data
There are many influences on color in the imaging process including lighting, optics, sensor, and monitor, and ultimately print. The first, and generally most important, is lighting. There are plenty…Read article -
3D Localization Microscopy With Ground State Depletion (GSD)
With the latest development of a GSD 3D super-resolution platform, it is now possible to achieve a lateral resolution of down to 20 nm and an axial resolution of 70 nm. The technology is based on an…Read article -
Step by Step Guide to Fluorescence Microscopy
Fluorescence Microscopy is a special form of light microscopy. It uses fluorescence to highlight structures in fixed and living biological specimens instead of using absorption, phase or interference…Read article -
Abstracts of the 3rd European Super-Resolution User-Club Meeting
The 3rd meeting of the Leica Super-Resolution User Club was held from June 17th to 19th, 2013 in collaboration with Alberto Diaspro and the Italian Institute of Technology (IIT) in Genoa. Confocal and…Read article -
The Principles of Polarization Contrast
Polarization contrast microscopy is a convenient way to make birefringent crystalline structures like starch grains or cellulose visible without staining. This tutorial will explain the optical…Read article -
50 Years of Image Analysis
Modern image analysis systems perform highly sophisticated image processing functions on images from an automated microscope and digital camera. 50 years ago, the first image analysis system was…Read article -
Abstracts of the 2nd European Super-Resolution User-Club Meeting
The 2nd meeting of the Leica Super-resolution User club was held from September 25 to 27, 2012 in collaboration with the Science for Life Laboratory at the Karolinska Institute, Stockholm, Sweden. …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 -
Widefield Super-Resolution with GSDIM
Great advancements in biology have been possible by using fluorescence microscopy. So far, the resolution of the images was limited due to physical constraints. In the past couple of years, new…Read article -
Image Processing for Widefield Microscopy
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…Read article -
The Principles of Modulation Contrast
Modulation contrast creates 3D-like images of unstained specimens, rendering it the contrasting method of choice for applications like e.g. in vitro fertilization, where DIC is not possible (due to…Read article -
Fluorescent Dyes
A basic principle in fluorescence microscopy is the highly specific visualization of cellular components with the help of a fluorescing agent. This can be a fluorescing protein – for example GFP –…Read article -
The Principles of Phase Contrast
In this tutorial the principle of phase contrast imaging is described taking the example of an inverted research microscope. Additionally, the alignment of the components needed for phase contrast is…Read article