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Microscopy and Imaging

Importance of Color Reproduction in Microscopy Images

Through our eyes, seeing is not always believing. Under different lighting conditions, we tend to see the same objects as having the same color. For example, an apple will appear red whether it is lit by daylight or candlelight and a white sheet of paper will be perceived as being white regardless of the light…

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How to Become a Live Cell Paparazzo: A Beginner’s Guide

Think of the very first time you looked at cells under a table-top microscope. Here’s what you would have done in that experiment: Step 1: Grow cells. Step 2: Plate cells on to a glass/quartz slide. Step 3: Insert the slide under a microscope and look. The protocol for performing single-cell microscopy has a similar…

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Introduction to Optical Fibers, Part 1

Everybody’s talking about them, tens of millions use them, but do you know anything about optical fibers/cables? Well you should. In this first (of many) articles I will cover the basics of optical fibers, what they look like and what they are made of. Optical Fiber Anatomy First thing first, the terms “optical fiber” and…

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Top Tips for Yeast Microscopy

Microscopy is one of the fun parts of working with yeast. If you fix your cells, you can get a snapshot of the structures. Live cells microscopy using fluorescent proteins tagged proteins is even better, as you can see the dynamics and cell machinery working before your own eyes. Light Microscopy to Check for Contamination…

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How to Troubleshoot Problems with Fluorescently Tagged Proteins

Using fluorescent proteins as imaging probes is a widespread and versatile technique in microscopy. You can use them in a wide range of living systems, from single cultured cells to complete organisms and animals. Fluorescently tagged proteins can be used to track and examine real-time localization, interactions and translocation of your protein of interest, as…

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The Power of STED Microscopy, Part 2: Sample Preparation.

In part 1, “The Power of STED Microscopy: How Does it Work?” I covered the basics of STED Microscopy: how it works and why you might want to use it. If you now want to do STED Microscopy, the next step is to optimize your sample. In this article, I cover how you can get…

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Live-Cell Imaging: Choosing the Right Technique

If you want to see in real time what is going on inside your cell then you should be performing live-cell imaging. Live-cell imaging techniques allow real-time examination of almost every aspect of cellular function under normal and experimental conditions. With all live-cell imaging experiments, the main challenges are to keep your cells alive and healthy…

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Catch of the Day: A Look into Different FISH Techniques

You are probably familiar with fluorescent in situ hybridization (FISH) to detect and localize the presence or absence of specific DNA sequences on chromosomes. But did you know there are numerous FISH experiment variations? Including high-resolution FISH and quantitative FISH? Read here about Fiber-FISH, Q-FISH, and Flow-FISH and decide if you would like to undertake one of…

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Slides 101

Would you eat your spaghetti dinner without a plate? No, of course not! It would make a big mess and be ugly to look at. Instead you NEED something to put that spaghetti on, to contain it, to keep it clean, to make it look nice – a bowl, a plate, SOMETHING! By the same…

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How to Measure FRET

Discover the different approaches to FRET quantification and get tips for selecting your FRET pairs.

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Acid Wash, Autoclave, Flame or Coat? Slide Basics Explained

There are about as many protocols to prepare coverslips as there are ways to make tuna casserole. You can spend from 5 seconds to 2 days, depending on what your lab prefers. But in the end, what’s really needed? I’ve tried many protocols over the years and I’ve questioned some steps. Here I share my…

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Vitamin H and Egg White: Streptavidin-Biotin for Immunohistochemistry

If you want to make molecules stick together you need to know about streptavidin/biotin. This article follows on from Mike’s article looking at ‘sandwich’ and ‘amplification’ methods of immunohistochemistry (IHC) and covers how streptavidin-biotin works in IHC, including protocols. Streptavidin-Biotin What is it? Avidin is a natural biotin-binding protein found in egg whites. Streptavidin is similar…

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PIER, HIER and Mannich: Antigen Retrieval in Immunohistochemistry

When you fix your tissue samples with paraformaldehyde (PFA) the proteins in your sample become covalently cross-linked. This is good to preserve the ‘architecture’ of your tissue sample. However, this cross-linking can become a problem when you carry out immunohistochemistry (IHC). Cross-linking can ‘mask’ or hide your antigens-of-interest and make them ‘invisible’ to your IHC…

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Microscope maintenance: changing and aligning your mercury burner

Mercury burners are one of the most common light sources used in fluorescence microscopes, producing a wide spectrum of wavelengths making them a great light source for viewing your samples. But they do have their issues – one of which is their propensity to break or become damaged if not used and cared for correctly, which…

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The power of STED microscopy, Part 1: How does it work?

Do you suspect that your favourite protein is doing something really cool? But you cannot see it because your confocal microscope’s resolution is limited. Then Stimulated Emission Depletion (STED) microscopy is what you need! With the power to smash through the diffraction limit of confocal microscopy, STED opens up a whole new world of improved…

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Is There Overlap? Find Out with a Proximity Ligation Assay

Colocalization blues (and reds and greens) Trying to find if and where two epitopes co-localize (or, to be more precise, where they are found in close proximity) may seem easy at first: 1) Bind your two epitopes with primary antibodies from two different species, 2) bind these primary antibodies with two secondary fluorescent antibodies, one…

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Digital Pathology – why you need it and how to choose the best camera for it

Yup, we really are in the digital age…even the pathology is digital. Facebook, Instagram, Tumblr. It has never been easier to take pictures and share them. The digital revolution is upon us and nothing is safe, even your pathology samples. Digitizing your pathology samples can help you better organize and manage pathology for later. And…

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Through the Looking Glass: Picking the Right Microplate Reader

You are finished with your super-arduous experiment. Now the fun part! To “read” your results with a microplate reader, but do you know which microplate reader to use? Or even that there are different types? Have no fear. I am here to help ye, scientific one! Read on for my pointers on picking the right…

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Looking good! A Guide to Adjusting and Maintaining Microscope Eyepieces

The magnification and viewing of samples using a microscope relies on both the objectives and the eyepieces working harmoniously together. If you buy a ready-to-use microscope, then the objectives and the eyepieces which are fitted as standard will be designed to complement each other. On the other hand, if you are designing and building a…

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The Who’s Who of Super Resolution Microscopy, Part 1

Super resolution microscopy shattered the lower resolution limit offered by traditional confocal microscopy, enabling us to see more detail. Discover the different super resolution techniques, how they work, and what they are being used for.

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I Can See (See Dee)! CCD and CMOS Cameras for Microscopy

Two different sensors are generally used in cameras for microscopy: Charge Coupled Devices (CCD) or Complementary Metal Oxide Semiconductors (CMOS or sCMOS). Although there are a number of similarities between the two sensors, differences in the way they function can have an effect on image capture time as well as signal-to-noise ratio. Let’s take a…

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Two Photon Confocal Microscopy: What it is and How to Use it to Your Advantage

“A two photon microscope has higher sensitivity than a normal confocal microscope, because it uses two photos instead of one!”  Yes, I can bear witness that this phrase has actually been uttered, and it was not by an undergraduate student. No exception to the rule The condensation of various levels of misunderstandings in this statement…

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Dots, Probes and Proteins: Fluorescent Labels for Microscopy and Imaging

If you remember from one of my previous articles (if not, you can read it here!), we introduced ‘fluorophores’. These are basically substances (natural or synthetic) which have the ability to absorb light at a low wavelength and re-emit at a higher wavelength. In other words- they fluoresce! In this article, I’ll introduce the three…

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“Where the hell did my signal go?” AKA The Problems (and Uses) of ‘Photobleaching’ in Microscopy and Imaging

Like most things in this world, fluorophores are mortal, and eventually your once bright fluorescent image will inevitably fade to black. This fading or ‘photobleaching’ of fluorescent signal can make imaging difficult, especially if you are trying to take quantitative images. Read below to learn what causes photobleaching of your fluorophores and how best to…

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Welcome to the Dark Side…Dark Field Microscopy That Is!

We all know that stars are easier to see against the dark background of the night than they are to see against the bright sky of day. Well did you also know that the same may be true of your microscope specimen? Dark field microscopy is a popular microscope technique that makes your unstained transparent…

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The Many Flavors of Widefield Microscopy

While confocal microscopy seems to have become pervasive in cell biology, widefield microscopy techniques still have a special and important place. This month on the Microscopy and Imaging Channel, we’re focusing on widefield microscopy techniques: covering the basics of what these techniques are and when you should turn to them. What is this ‘Widefield’ you…

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Catching Waves: What a Microscopist Ought to Know About Phase Contrast

Phase contrast microscopy is a light microscopy technique which is primarily used to visualise live cells. Using various filters and condensers, the image produced by phase contrast allows us to see greater detail in live cells and can highlight aspects such as intracellular structures. Keep your cells alive! The best way to view cells is…

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5 Digital Photomicroscopy Competitions

Have you ever considered that your data is a work of art in the making? In particular, microscopy has the potential to capture the clearest, most stunning moments in life. And when a scientist accomplishes that, it’s a feat to share with everyone.  Why not share it through a photomicroscopy competition? Photomicroscopy competitions quickly help…

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How to Make Sure You Never Lose a Lab Sample Again

It’s 11 PM…do you know where your samples are? If you dread the thought of having to hack through ice, snow and the bone-chilling depths of every freezer to locate them, flirt with frostbite only once. Better yet, never hunt for missing lab samples again! Here are some useful ways to keep track of every…

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