Toluidine Blue – A Histology Stain for Mast Cells (and Other Things!)
Discover the magic of toluidine blue – a polychromatic dye that changes color depending on which tissue component it is staining.
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Discover the magic of toluidine blue – a polychromatic dye that changes color depending on which tissue component it is staining.
Blunt-end cloning involves the ligation of DNA fragments – usually between a plasmid vector and an insert – whose terminal ends are not “sticky”. Performing these ligations is notoriously difficult, particularly with large DNA fragments. But it is possible. And in this article I’ll give you some tips that I hope will increase your chances…
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…
Get introduced to some of the special stains for histology and learn some top tips for getting great results.
If you’re picking up a list of available plasmid vectors for the first time, it can be mind-blowing to decide on the best one for your experiment. What cloning sites do you need? What restriction enzyme sites? What insert size is possible? Let me help you with some pointers about what factors to look out…
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…
Looking for paraffin alternatives for tissue embedding? Find out the benefits of cryo and resin tissue embedding and how they work.
The lac operon is an amazing tool in molecular biology. It has been used for decades to turn on protein expression in an inducible manner with IPTG. The result is synthesis of vast amounts of protein to be used as you wish. While the lac operon is an amazing tool for protein production, it is…
Removing proteins from acqueous solutions can be a pain if you have loads of samples to do. This article explains how to prepare the DIY phase separating gel to easily and effectively separate two-phase mixtures, saving you time and money.
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…
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…
Scientific creativity and repetitive tasks do not go well together. There is nothing worse than having to repeat the same task over and over again – I know somebody who quit science because of this and became a castle curator. So how do you minimise the amount of one of the most repetitive tasks –…
Multiplexing qPCR enables fast, precise DNA quantification by combining amplification and detection in one reaction. This advanced PCR technique conserves samples, reduces costs, and measures multiple targets simultaneously using fluorescent probes. With careful assay design and optimization, multiplexing qPCR improves efficiency and accuracy across molecular diagnostics, gene expression studies, and pathogen detection workflows.
Counterstaining can have a big impact on your histology result. This short guide will introduce you to some available counterstains providing you with a few more choices.
Achieving a good immunohistochemistry signal-to-noise ratio involves many factors, including a good blocking protocol. Read on to learn about blocking non-specific staining in IHC.
Plasmid mapping and DNA annotation software are pretty abundant these days, alongside tools for viewing and comparing raw sequence data. A quick Google search brings up dozens of hits – but how do you know which one to use? If you are like most molecular biologists, you probably use the same software your colleagues do—usually…
Haematoxylin and Eosin staining is the most common staining in the modern (and old!) histology lab. This staining technique gives an overview of the structure of the tissue and can be used in pathological diagnosis. This article follows on from Nicola’s introduction, but we’ll take an in-depth look at the stains, chemistry and method to…
The Irish Famine (or ‘Great Potato Famine’ if you live outside the Emerald Isle) killed one million people and forced another million to leave the country between 1845 and 1852. It was caused by a blight on the country’s main food stock- the Irish ‘Lumper’ potato. Now, researchers have identified the genome of the blight…
I was first introduced to Conrad Waddington’s epigenetic landscape when reading ‘The epigenetic revolution’, a fantastic introduction to epigenetics, and in my opinion, a must read for anyone who is looking for an entertaining and enjoyable introduction to this fascinating field. In his model, Waddington likens the process of cellular differentiation to a marble, which…
Unlike immortalized cells lines, primary cells can only be kept in cell culture for a finite period of time, if at all. Therefore, you often need to obtain primary cells directly from an animal source. After which, you may fix and image the primary cells in situ (as part of the whole organ), or you…
Isoelectric focusing electrophoresis (IEF) of proteins is nowhere near as popular as its cousin – sodium dodecyl sulphate-polyacrylamide gel electrophoresis aka SDS-PAGE. While in both methods the proteins are denatured, IEF is a gel-based electrophoretic separation of proteins using difference in their overall charges. The sodium dodecyl sulphate – SDS part of the usual gel…
Fixing suspension cells for imaging can be trickier than fixing adherent cells, as they can’t be cultured on a coverslip. Discover how you can stick them down with the help of centrifugation.
Wouldn’t it be great to put your nucleotide sequence into a program and get back a 3D-structure of your protein and a full description of its functions? In theory, because the protein 3D-structure is determined by the aminoacid sequence, given the right algorithm and a powerful enough computer, this should be simple. In practice, because…
You just can’t put raw tissue or cell samples on your slides and expect good histology results! Instead you must preserve or ‘fix’ your samples. Fixing ensures that your cell structures stay intact and that your antigens are immobilized. Ideally, fixation would also still permit unfettered access of your antibodies to your antigens. However, as…
According to the central dogma of molecular biology, DNA is transcribed into RNA, that is translated to proteins. Inconveniently, the vast majority of the genome contains sequences that do not actually code for proteins. So, this non-coding RNA (ncRNA) was dismissed as non-functional junk, letting researchers tick the box on their to-do lists and head off…
The theory behind the idea of having shared microscopes is a good one, but, in reality, this can sometimes mean you have to put up with the dirty habits of your fellow scientists and researchers. And some of your lab mates turn out to be really mucky! Here’s my Top 10 of things which really…
Recently BsB author Yevgeniy Grigoryev shared a total RNA isolation protocol. The one I use is even simpler—no expensive Trizol, which is a mix of phenol and some salts, all that is required is some Tris, SDS and phenol/chloroform mix. I have never used this protocol on non-yeast cells but I am almost sure that…
It is the black death of cell culture. Scientists don’t dare utter its name and many a graduate student has fallen victim to its indiscriminate menace. These stealthy anarchists infiltrate quietly but deliberately until their numbers swell and then they attack in strength, overwhelming their victims before they can put up a fight! What is…
Have you ever emerged from the lab, bleary-eyed, blinking dazedly at the sun after spending hours hunched over a lab bench, counting endless bacterial colonies or viral plaques? A necessary evil… I consider colony/plaque counting one of the necessary evils of working with microorganisms. Necessary because many experiments have an endpoint that requires determining the…
“Do we use a monolayer or 3-D cell culture for our experiments?” A simple, yet puzzling question asked by my group leader while I was working on a drug development project at the University of Abertay Dundee. How do you answer such a question? Just read on to find out! Monolayer vs spheroid One of…
The field of epigenetics is exploding and given the strong links between epigenetic state and disease, the need to study markers like DNA methylation in humans is very relevant. This article outlines some of the main factors you should be taking into account in your study of DNA methylation in human tissues. Here goes: Biological…
Did you know fixation can mask antigen sites in your sample? Discover how you can unmask them and get your signal back on track!
Whole genome sequencing (WGS) is becoming increasingly common. Doctors now routinely order it for patients with puzzling diseases. The NHS (National Health Service in the UK) has declared that it will sequence 100,000 genomes over the next few years. Increase WGS…increase ethical questions The direct-to-consumer company 23andme has been experimenting with whole exome sequencing (WES), and another company, DNA…
To answer some of the more interesting research questions, you often need to get a good look at what’s going on inside the cell. Whether you’re running a Western blot or measuring enzyme activity, many assays require access to the materials (e.g. proteins, DNA, subcellular fragments) contained within the cell walls. There are several ways…
As part of my job ensuring plasmid quality at Addgene, I analyze 50-100 sequencing reactions a week. So I have developed some good habits that I wanted to pass on to you to make sure you are getting the most out of the data you get back from your sequencing runs. The most important of…
You’ve been told that maintaining a sterile environment in a tissue culture hood is vital to preventing contamination of cell cultures. But what exactly is meant by sterile? The definition of sterile is ‘completely clean, sanitized, and free of all forms of life’. Obviously you still want your cells and/or any other organisms you are…
Have you ever isolated a great little population of cells after days or months of trying, got truly excited about doing some immunofluorescence with them only to find out (at the very end) that all your cells washed away?! If this has happened to you, then look no further; we will introduce you to some…
Most ‘wet lab’ biologists do not have much computer programming experience, which can make downstream analysis of next generation sequencing results a bit daunting. After the sequencing platform spits out your data, what do you do with it? That’s where Galaxy comes in. What is Galaxy? Galaxy is a bioinformatics workflow management system, created by collaboration…
Unlike DNA, which can last for eons, RNA is a fragile and degradation-prone cousin. After working with RNA for a while, one becomes quite paranoid about handling RNA because even a single sneeze or drop of saliva can potentially affect your results. The reason is that there are enzymes called RNases that specifically target and…
In the previous article on EC numbers, I explained how the Enzyme Commission names enzymes, and why it is so important. In this article I’d like to take you on a brief journey through the history of the Enzyme Commission. Like many histories in science (e.g. this!), it is fascinating and gives a useful perspective…

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