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Microscopy and Imaging

New 2D Superresolution mode for ZEISS Airyscan – Fast and gentle confocal imaging with 120 nm resolution

New 2D Superresolution mode for ZEISS Airyscan – Fast and gentle confocal imaging with 120 nm resolution Speakers Joseph Huff, PhD Tutorial abstract Utilizing a pinhole-plane imaging concept, ZEISS Airyscan allows for simultaneous improvement in resolution and signal-to-noise by capitalizing on an innovative 32-channel GaAsP photomultiplier tube (PMT) array detector. Each detection channel functions as…

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How to Conduct Localized Proteomics of Microscopic Regions

Precise and lightning-fast spectral Fluorescence Lifetime Imaging at video rate integrated in a high-end confocal microscope In this tutorial, you will learn: How to perform isolated proteomics of microscopic regions of interest How to use this technique with FFPE tissue How proteomics can help you understand molecular mechanisms in disease Speakers Dr. Eleanor Drummond Tutorial abstract…

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From Cells to Scope: Chamber Slide Immunochemistry

Immunolabeling is the tried-and-true immunochemistry method of getting the stain you want onto the molecular target you want. Whether that target is contained within a large region of tissue (immunohistochemistry) or inside a single cell (immunocytochemistry), the ability to accurately label large numbers of samples will simplify your workflow and help you to achieve excellent…

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Lasers for Confocal Microscopy: Part II

Continuing from our first article on lasers for confocal microscopy, we will now discuss two specialized types of lasers: lasers for two-photon excitation and tunable, white light lasers. We will also discuss the applications of the two lasers. Lasers for Two-Photon Excitation The two-photon absorption phenomenon was first described for microscopy in 1931. Here, the…

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Things to Consider When Buying a Microscope Camera (Part 2)

In the first part of this series, we discussed the differences between a color and a monochrome microscope camera and when one is advantageous over the other. We also touched on the subject of optimal camera resolution for a given imaging system. In this part, we will tackle a few additional camera specifications and how…

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Biological Applications of X-Ray Microscopy and Correlative XRM – FIB-SEM Imaging

In this webinar, you will learn about the power of 3D imaging of larger samples.  In particular, you will: Gain insight in to how to use x-ray microscopy to analyze biological samples Learn the basics about sample preparation for x-ray microscopy Understand the benefits and drawbacks to different imaging conditions X-Ray Microscopy (XRM) is a…

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Large Volume Serial Blockface Electron Microscopy Imaging

Large Volume Serial Blockface Electron Microscopy Imaging An unlikely renaissance is occurring in biological imaging, propelled by the ongoing development of serial blockface electron microscopy imaging technologies. Imaging systems based on scanning EM instruments place large-volume automated EM serial imaging within reach of most labs.  How best to take advantage of the automation and versatility?…

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Things to Consider When Buying a Microscope Camera (Part 1)

Purchasing a microscope camera is one of the most daunting tasks you might have to undertake. Before you set out to buy that camera, carefully consider your applications. Things like sample brightness or the speed of the phenomenon you are trying to capture can dictate your choices. Also, this is the time to make peace…

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All You Need to Know (and More) About Embedding for Electron Microscopy

Interested in the detailed structure of your tissue? High-resolution imaging techniques, such as brain electron microscopy, provide an intricate view of your tissue. While it may be a rather complicated procedure with nasty chemicals, the advantages of epon embedding can make it the best choice for morphological studies. The hard blocks are excellent for structural…

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Tissue Embedding Throwdown: Paraffin vs OCT vs Resin

Tissue embedding and sectioning is a backbone of many biological research labs. While commiserating with other grad students over tedious hours spent in the lab, you’re probably aware that there is more than one way to slice up a chunk of tissue. We’ve previously introduced what to consider when choosing a tissue embedding medium and…

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How to Use CellProfiler for Cell Imaging

Are you trying to figure out how to calculate intensities of fluorescently-labeled single cells? Do you have cells at high densities or present in clusters? Are you worried that your current cell imaging analysis software is unable to mark clear boundaries around each cell in a cell cluster? Don’t fear, because CellProfiler 2.1 is here to…

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Optocardiography: Optical Imaging of Cardiac Physiology

In this webinar, you will learn how to use optical imaging for functional cardiac physiological mapping of transmembrane potential, calcium handling, and metabolism. The main aspects covered in the webinar include: How to design an effective optocardiography system and select optimal fluorescence probes for your research needs using open source hardware and software. Troubleshooting tips…

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Unpacking the Daunting Task of Stereology for Electron Microscopy

Unpacking the Daunting Task of Stereology for Electron Microscopy Electron microscopy provides fantastic detail and resolution, with brain electron microscopy allowing visualisation of neurons and their individual synaptic connections. You may find yourself needing to count these neurons or connections, which can easily go into the billions. But counting these one-by-one isn’t really feasible. This…

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Detecting Cell Apoptosis on Tissue Slides

Apoptosis, or programmed cell death, is a physiological process in which individual cells are set to die without harming their environment. It involves a cascade of complex and tightly regulated cellular events. Detecting apoptosis on tissue slides, involves either detecting the molecular participants of these events, or the morphological changes that occur on the cellular…

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Mouse Whole-Brain Volume Electron Microscopy for Cellular Connectomics – Enabling Large-Scale SEM Projects with ZEISS MultiSEM

Recent advances in high-throughput multi-beam scanning electron microscopy (EM) and mouse whole-brain EM preparation and collection on tape (“Brain-on-Tape”) have resulted in substantial progress towards a nano-scale map of the whole mouse brain. These maps can be used to determine how individual neurons are synaptically connected and can be used to reconstruct the precise “wiring…

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Enhance Your Imaging with Super-Resolution: Improve Spatial Resolution and Increase Signal to Noise Ratio in Difficult Samples

In these informative case studies, you will learn how: HyVolution 2 can improve the spatial resolution you get from your sample, whilst remaining gentle to the fluorophores HyVolution 2 overcomes issues experienced with other imaging modalities, even in difficult samples such as plant material, increasing resolution and signal to noise ratio, thereby facilitating downstream image…

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Harnessing the Power of Heterogeneity in Cryo-electron Microscopy

A major impediment in the determination of high resolution protein structures by single particle cryo-electron microscopy has been the presence of sample heterogeneity. Oftentimes, heterogeneity is due the dynamic nature of protein complexes, which can exist in multiple different conformational states in solution. Recent advancements in cryo-electron image processing have provided tools to accurately sort…

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Automated Microscopy

The traditional microscope that you know and love is operated manually. Picture the scene: the microscopist chooses the light source, gently places the sample the moveable stage, selects the objective lens, and scans to select the field of view. This process is perfect for processing and analyzing a small number of samples per day. But…

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Revealing Cellular Dynamics with Millisecond Precision – The New Tool That Turned Electron Micrographs into Motion Pictures of Neural Communication

Revealing Cellular Dynamics with Millisecond Precision – The New Tool That Turned Electron Micrographs into Motion Pictures of Neural Communication In this tutorial: What if you can dissect the cellular dynamics with millisecond precision? What if you can unravel the morphological transformation of a neuron millisecond by millisecond using electron microscopy? Could this be even…

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In Situ Zymography: Let’s Catch that Enzyme in Action!

In situ zymography (ISZ) is the best choice to study proteases. Proteases are a challenge to study as proteases are extremely potent enzymes. As such, they need to be controlled at multiple levels to prevent them from being unleashed and making a cellular mess. Regulation of their activity occurs at virtually all levels: transcriptional control,…

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Automated Cell Counting with a Fluorescent Twist

Cell counting is the bane of existence of many researchers. Countless hours spent in front of the microscope with a haemocytometer on the stand and a manual tally (or “clicker”) in hand can be really daunting. Not to mention that no one will ever double check your count if you don’t take a picture. Those…

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Analyze Immunostained Slides with Semiquantitative Scoring

A  routine task in the lab is to investigate the presence of your favorite protein in a range of histological samples. No doubt, staining your tissue sections using good old immunohistochemistry (IHC) would be your first choice. You just got to love a technique that has celebrated its 70th birthday, and is still used in…

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Tips for Peering into the Interior of Mice Using Intravital Microscopy

Techniques to study entire tissues, such as brain imaging microscopy, provide great insight into the biology of the whole tissue, rather than just individual cells.  Taking this one step further is intravital microscopy (IVM); a newer approach for the imaging of living tissues and organs in live animals. A wide variety of organs can be…

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Data Analysis for Three-dimensional Volume Scanning Electron Microscopy

In recent years, three-dimensional (3D) scanning electron microscopy techniques have gained recognition in the biological sciences. In particular, array tomography, serial block face scanning electron microscopy (SBFSEM) and focused ion beam scanning electron microscopy (FIBSEM) (described in Three-Dimensional Scanning Electron Microscopy for Biology) have shown an increase in biological applications, elucidating ultrastructural details of cells…

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Microscopy – a Numbers Game

While the microscope is synonymous with biology, it is a child of physics and technology. When we learn about the microscope we learn physics—specifically, we learn about optics. Many great resources are available that explain the inner working of microscopy. And, like most things in physics, the inner working of microscopes comes down to a numbers…

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The Art and Science of Figure Creation: Think BIG to see Small

There are those of us who began our careers literally in the dark. Yes, there was a time and not that long ago, that all figures had to be on film. Slide presentations were slides. Micrographs were, well, micrographs on film. Figure creation involved several steps: figures for publications had to be mocked up; then…

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Halogen vs LED Lighting in Digital Microscopy

When it comes to light sources for microscopy, there is really no such thing as the “best.” The type of light source you use depends on the system you are working with and the type of result that you want. Digital systems are usually designed to work with either a halogen or a LED light…

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Challenges of Autofluorescence in Neuroscience

If you have ever imaged biological samples, you have likely encountered autofluorescence. That pesky background coloration you see under the microscope, which can make it difficult to distinguish your actual signal from the noise.1 When you are trying to look for something as delicate as RNA, you don’t want to be hunting for your signal…

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How to Maintain Live Cells on a Microscope Stage

Are you preparing to set up live cell imaging experiments? You’ve got all your cell lines, antibodies, reagents, and protocol ready. You just want to wake up in the morning and enter into that dark room. Well, think again!! As we (I mean the cell biologists) always say, happy cells mean happy life. You have…

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How to Quantify Images in an Unbiased Way

Microscopists like to say that seeing is believing. The so-called belief in this context is a product of highly experienced imagination, which raises a concern about the objectivity. To remain objective, we try to use analytical approaches to quantify images and turn phenotypes into numbers. Many image analysis approaches exist nowadays, from simple manual measurements…

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Controlling Color Image Quality in Microscopy: Start at the Beginning

The only constant with microscopy imaging is variability in both color and image quality. You only need to look at images in journal articles, posters, around your laboratory, or compare your images with a colleague’s—the evidence is staggering. Interestingly, variability doesn’t generally come from the digital camera, rather it comes from our use of imaging…

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Quick and Easy Automatic Cell Counting

Are you wondering how on earth you’re going to count thousands of cells across a stack of images? Well, I’m going to show you a simple method for automatic cell counting with ImageJ. For those of you unfamiliar with ImageJ, it’s a popular image processing program that runs on Mac, Windows, and Linux. Assuming you…

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Three-Dimensional Scanning Electron Microscopy for Biology

Scanning electron microscopy (SEM) is a powerful technique, traditionally used for imaging the surface of cells, tissues and whole multicellular organisms (see An Introduction to Electron Microscopy for Biologists)(Fig. 1). While the resultant images appear to be three dimensional (3D), they actually contain no depth information. However, there are several SEM techniques that can obtain…

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Color Image Quality Control in Microscopy

In this webinar, you will learn about color reproduction in microscopy images—and how to fix it if your color reproduction goes awry. The main points we will cover are: The reasons that could color reproduction go wrong in your microscopy images How to correct your microscopy images with inappropriate color reproduction How to be ethical…

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