Articles by mikemillar:

Can’t See the Mouse for the Mice?! Solutions to Mouse-on-Mouse Immunohistochemistry Detection

The issue of mouse-on-mouse background is only a cause for concern for the histotechnologist working within a research environment. Those working in a diagnostic setting will probably never experience this as they will be working with human tissue with antibodies raised in a variety of species- but one species that won’t be used is human!…

18 Mar 2014 Microscopy & Imaging

Fluorescent Tyramides In Histology: A Versatile Approach For Multiplex Molecular Tissue Probing.

If you are a researcher who is using fluorescent staining on your cells and tissue sections, you should seriously consider using tyramide reagents. Why? Well, because they can have exquisite sensitivity, are easy to use, versatile and there is an increasing range of fluorophores in the pipeline. As they rely on a peroxidase catalysed visualization…

21 Aug 2013 Microscopy & Imaging

Immunohistochemistry- PAP, APPAP and Sandwiches!

You’ll give me an (enzymatic) complex! Following on from Part 1 of this article, let’s start by having a look at the two most popular enzymatic ‘sandwich’ methods; The Peroxidase anti Peroxidase method (PAP). The PAP method was the first sandwich method that I used and involves three main stages- application of primary antibody, secondary…

12 Feb 2013 Microscopy & Imaging

Immunohistochemistry- Direct vs. Indirect Methods, and a Golden Rule

In my previous article I covered different immunohistochemical staining techniques at a superficial level. In the following articles I will start to explain these technologies in a bit more detail and in which situations they should be applied. All of the following will involve additional stages when applying them, for example- serum blocking, protein blocking,…

29 Jan 2013 Microscopy & Imaging

Immunohistochemistry- an Introduction, Techniques and an Evolution Towards Robots!

The last two decades have seen a dramatic increase in the number of publications using immunohistochemistry (IHC) as a research tool to identify the spatial location of proteins of interest within cells, tissue sections and whole-mount preparations. Grinding and binding The advantages over ‘grind and bind’ methods are apparent, but the very best results will…

15 Jan 2013 Microscopy & Imaging

Tissue Embedding and Sectioning: Something to Think About Whilst in the Bath.

In the same way that you should ‘Think Before You Fix’, the choice of embedding media should be dictated by your required end-point. The basic principle is that by processing tissue into an embedding medium you harden the tissue and provide support protecting it from the mechanical forces associated with sectioning. Parma ham and steak…

16 Oct 2012 Microscopy & Imaging

An Introduction To Fixation For Histology: Think Before You Fix!

For many cell or tissue based experiments, microscopic imaging is likely to be  the end point, and  in this series of articles we want to illustrate how some joined-up thinking regarding tissue or cell collection, fixation and processing, subsequent staining, microscopic imaging and analysis will produce better results. Make imaging easier For example, intense well-stained…

17 Jul 2012 Microscopy & Imaging