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Displaying posts in the category: How to

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

How to use Dragonfly Spinning Disk Microscope for Multiplex In Situ Hybridization

Sponsored by Andor, on 4 November 2020

Multiplexing in cell biology is the unveiling of several (Xn) RNAs in its 2D or 3D biological context. Multiplexing has become a hot topic in neurosciences, oncobiology, disease target diagnostics, development, behavioural studies, etc. Several techniques have been developed to allow multiplex imaging; examples of such are FISSEQ, instaSEQ, osmFISH, STARmap, MERFISH and seqFISH. The advantage of

Primers on Microscopy for Biologists - Resolution

Posted by , on 15 July 2020

Formal definitions of resolution refer to imaginary objects such as infinitely small sources of light. I will avoid those and instead try to provide a pragmatic explanation. Practically, the spatial resolution is the size of the smallest structure that can be distinguished in the light coming from a specimen. All sensors and components of the

How CLIJ2 can make your bio-image analysis workflows incredibly fast

Posted by , on 14 July 2020

Do you also spend a substantial amount of your time waiting for automated image analysis to finish? I did, again and again for more than a decade. And, then,  the morning after running a script overnight, you realize that a parameter was wrongly set. It was about time to change that. History Processing on graphics