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Displaying posts in the category: How to

Using an autocollimator to align 4f systems

Posted by , on 29 June 2022

Graeme Johnstone, Brian Patton, Department of Physics and SUPA, University of Strathclyde, Glasgow G4 0NG, United Kingdom Modern microscope systems comprise many optical elements with precise alignment tolerances. As such, as the complexity of the system increases, the challenge to align them rapidly becomes a major hurdle to implementing home-built designs. One recurring problem is

If you can detect it, you can track it - recent TrackMate developments

Posted by , on 17 February 2022

TrackMate is your buddy for your everyday tracking  TrackMate (Tinevez et al., 2017) is an open-source Fiji plugin initially made for tracking single particles imaged with fluorescence microscopes. It has now been updated to include a collection of new detectors and direct implementation of deep-learning models (Ershov, Phan, Pylvänäinen, Rigaud et al., 2021). In addition

Protein micropatterning: beauty standards in cell culture

Posted by , on 16 February 2022

When imaging cells grown on a flat substrate, such as a glass coverslip, we quickly admire the diversity of morphologies different cells can take on. Some cells, such as COS7, will take on a flat “pancake”-like shape, which makes it easy to image its organellar structure in a 2D plane. On the other hand, a

Janelia+EMBL BioImaging Seminar Series: How We Started a Successful Seminar Series during the Pandemic

Posted by , on 25 January 2022

How we started a global BioImaging seminar series in the middle of a global pandemic...

Behind the Screen - behind the scenes of setting up a dynamic screening platform

Posted by , on 22 November 2021

The Cell Biophysics lab has focused on FRET (Forster Resonance Energy Transfer) as it is a powerful technique to investigate dynamic protein-protein interactions, along with studying almost every aspect of cellular signaling with biosensors. FRET is detected either by ratioing the  intensities of the FRET donor and acceptor,  or by FLIM (Fluorescence Lifetime Imaging ;

PUMA Open Source Multimodality 3D Printed Microscope

Posted by , on 26 October 2021

PUMA is an advanced direct vision 3D printed multimodality microscopy system with fluorescence ,phase contrast, Köhler illumination and augmented reality functions - and more.

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.