People holding hands to represent interactions that can be uncovered using co-immunoprecipitation

Co-immunoprecipitation Protocol: Your Practical Guide To Co-IPs

Do you wonder if your favorite protein interacts with another protein? Do you wish that you could shine a spotlight on your protein to determine its binding partner? You can use co-immunoprecipitation (Co-IP) to find your protein’s partner. This article will get you ready for your first Co-IP, provide a handy Co-IP protocol, and discuss…

Let’s Talk About Stats: Comparing Multiple Datasets

Let’s Talk About Stats: Comparing Multiple Datasets

Last week I focused on the left-hand side of this diagram and talked about statistical tests for comparing only two datasets.  Unfortunately, many experiments are more complicated and have three or more datasets.  Different statistical tests are used for comparing multiple data sets. Today I will focus on the right side of the diagram and…

Fluorescence 101: A Beginners Guide to Excitation/Emission, Stokes Shift, Jablonski and More!
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Fluorescence 101: A Beginners Guide to Excitation/Emission, Stokes Shift, Jablonski and More!

You may already use fluorescence as a tool in your microscopy and imaging work, but, do you know exactly what it is? Why are certain proteins and probes fluorescent? What causes this light emitting property? We’ll have a look at these and more questions in this article. Start with a definition We’ll start with a…

Ten Tips for Turning Beastly Western Blots Beautiful

Ten Tips for Turning Beastly Western Blots Beautiful

Western blots can be ugly. I mean down-right, horrifically, wall-of-shame ugly.  Not only can they be embarrassing to show to your colleagues, but the ugliness can obscure your results, making it impossible to interpret your data. Blotting consists of many experimental steps, which makes the technique naturally error-prone. Although standardized protocols exist, many fail to…

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

“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…

Let me introduce you to ELISA…No, not the girl…The assay.

An ELISA (Enzyme-Linked ImmunoSorbant Assay) is a popular assay that uses antibodies and color change to detect proteins, peptides, antibodies or biomolecules in complex mixtures. ELISAs are popular because they are reliable, specific, easy to use, and can easily be scaled up to process multiple samples simultaneously. How an ELISA is Done: In an ELISA,…

Gibson Assembly: an easy way of molecular cloning!

Gibson Assembly: an easy way of molecular cloning!

Every hard-core biologist knows designing the perfect construct can be a complex puzzle to solve. This challenge, if successful, can be extremely satisfying but can also drive you crazy for weeks. Luckily, Dr. Daniel Gibson and his colleagues at the J. Craig Venter Institute designed a new easy-to-use cloning method. Better yet, this system allows…

Light Through Crystals: What Exactly is Differential Interference Contrast Microscopy?

Light Through Crystals: What Exactly is Differential Interference Contrast Microscopy?

Although his name could fit in easily to the early 1980’s Hip-Hop Scene, Jerzy Nomarski (or ‘George’) was actually a Polish physicist with an interest in optical theory. Born in 1919, he eventually became a member of the Polish Resistance fighting in the Second World War. He was captured by enemy forces and held as…

The Many Flavors of Widefield Microscopy

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…

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

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…

Tower of Babel: Next Generation Sequencing Provides New Insights on Chromosome Construction

Tower of Babel: Next Generation Sequencing Provides New Insights on Chromosome Construction

Biologists have long appreciated the complexity of genome organization, but until recently lacked the tools to discern the intricacies of this puzzle. Now, thanks to some handy cross-linking, careful amplification, and (of course!) next generation sequencing, teams from Massachusetts are taking us down the rabbit hole, with some surprising findings from Wonderland. Bend Over Backwards…

What Makes a “Good” Laboratory Buffer?

Just about any molecular biology experiment will involve the action of enzymes or other active proteins. And when enzymes are involved, the pH of your experimental environment is going to change. This is because most enzymatic reactions involve the loss or gain of hydrogen ions (protons), which modifies the pH of the environment. Biological systems…

How RNA-FISH Can Complement qPCR, and Vice Versa
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How RNA-FISH Can Complement qPCR, and Vice Versa

In studies of RNA abundance and gene expression, no one technique can answer all of the questions that need to be asked. So it is necessary to use a variety of experimental methods in concert. Two RNA detection and measurement techniques that complement each other well for this purpose are RNA Fluorescence in situ hybridization…

Never Leave an Empty Seat – A Simple Rule to Increase Your Networking at Scientific Talks

Never Leave an Empty Seat – A Simple Rule to Increase Your Networking at Scientific Talks

One of the great things about being scientists is that we are lifelong learners.  Scientists go to a lot of seminars, meetings and conferences.  The traditional, hour-long seminar is also a great way to learn a lot in a relatively short time. So it continues to amaze me how most scientists don’t take advantage of…

A primer on Probe-based SNP genotyping
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A primer on Probe-based SNP genotyping

Single Nucleotide polymorphisms (SNPs), colloquially pronounced ‘snips’, are the most common type of genetic variation in people. By definition, a SNP represents a single nucleotide variation at a specific location in the genome that is found in more than 1% in the population. For example, a SNP can replace the nucleotide cytosine (C) with an…

This One’s Upside Down! Inverted and Stereo Microscopes in Bioscience Laboratories

This One’s Upside Down! Inverted and Stereo Microscopes in Bioscience Laboratories

Most of the microscopes you will encounter in your laboratories will be ‘upright’. In other words, they are assembled (from top to bottom) in the order of; eyepieces, objectives (on revolving nosepiece), stage, sub-stage condenser, diaphragm and base. However, there are two other types of light microscopes you will perhaps encounter (and use) and it…