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Displaying posts in the category: How to

Using Nile Blue and Nile Red to visualise the presence of lipids in cryosectioned tissues

Posted by , on 14 September 2021

Lipids are crucial elements of mammalian (and non-mammalian) cell biology and yet lipids are challenging to visualise in situ. In comparison to proteins, which we can generate antibodies for, or carbohydrates, some of which we can detect using fluorescent lectins, there are relatively few lipid-specific fluorescent probes. Many lipids are highly conserved across species making

Temperature control in light microscopy – challenges and solutions

Sponsored by Interherence, on 9 September 2021

We describe common challenges that researchers face when trying to precisely control the temperature of the sample they are imaging. We aim to show that thermal control in high sensitivity microscopy is not trivial if you care about temperature directly in the field of view of the microscope (which you should). We hope to persuade

Collaborative bio-image analysis script editing with git

Posted by , on 4 September 2021

TL;DR: I’m a computer scientist who often collaborates with biologists on bio-image analysis scripts. We are using more and more git, a version control program, for working on code collaboratively. When using git, we speak about repositories, commits and pushing to the origin. We also make forks, send pull-requests and merge code. This blog post

Considerations for expression of fluorescent proteins and imaging in mammalian cells

Posted by , on 7 July 2021

Introduction to fluorescent proteins  Fluorescent proteins have the property of absorbing light at one wavelength and emit light in a longer wavelength. These proteins were observed first in bioluminescent organisms known to humanity for centuries. We can find examples of light-emitting organisms in multiple taxa: from single cell organisms like bacteria, to vertebrates like fish.

A First Exposure to Super-Resolution Microscopy

Posted by , on 11 June 2021

Biomedical research encompasses several fields of expertise involving complex biological topics and technologies. Studying a given subject is a process that takes years, decades, and sometimes a lifetime to complete. Consequently, researchers tend to become highly familiar with a specific subset of scientific topics and experimental approaches. However, they are often confronted with the cumbersome

Preparing your manuscript: guidelines for writing microscopy methods and figures

Posted by , on 25 May 2021

A new paper has caught your eye on Twitter or Pubmed based on the title. Next step? A quick look at the abstract. Still interesting? Let’s have a look at the figures. This is probably the most important element of the paper to attract the interest of the reader. If figures are easy to interpret

Phototoxicity - the good, the bad and the quantified.

Posted by , on 14 May 2021

Our virtual meeting on phototoxicity was held in late January 2021, generously sponsored by the European Microscopy Society and enabled by the Royal Microscopical Society. In four hours, spread over two days, the five organisers and twenty invited participants discussed the problem of phototoxicity in live imaging, and how we can start to tackle this

Part V The future: The hope of smart microscopes and phantoms

Posted by , on 24 April 2021

Elisabeth Kugler 1 and Emmanuel G. Reynaud 2 Contact: kugler.elisabeth@gmail.com; emmanuel.reynaud@ucd.ie We are reaching the end of our LSFM journey. Now it is time to look into the future, riding the plane of light! So, what is next for the LSFM? 1. Four-to-two, then four, then one. The story of SPIM, and LSFM per se,

A biologist’s checklist for calcium imaging and optogenetic analysis

Posted by , on 12 April 2021

Technological advancement constantly makes these methods more accessible, however, there are a number of understated complexities involved with these types of imaging-based experiments

LSFM series – Part III: Image acquisition: Calibration and acquisition

Posted by , on 17 December 2020

Elisabeth Kugler 1 and Emmanuel G. Reynaud 2 Contact:kugler.elisabeth@gmail.com; emmanuel.reynaud@ucd.ie Now, we are getting closer to the point of setting up our sample in the chamber to image it. Now that we know which microscope we are about to use (Part I) and have mounted it the right way (Part II), we need to trust