Newcastle University

Two dimensional (2D) images of fixed specimens can yield a lot of information and indeed have been the mainstay of the Bio-Imaging Unit for some time.  However, gone are the days where this imaging approach can lead to cutting edge science, in many cases there is a need to put these 2D images into the context of the live-cell environment.  To do this you need the ability to view cellular events as they happen when they happen and in real time.  The Bio-Imaging Unit with the support of the Faculty of Medicine has agreed the purchase of two advanced live-cell imaging systems in addition to upgrading the live-cell capabilities of existing ones.  With this, it is hoped that an extra dimension can be added to the Faculty’s cellular imaging capabilities.

Nikon Hybrid confocal microscope

The purchase of two state of the art Nikon A1R hybrid confocals brings with them access to hitherto unavailable live-cell imaging techniques while at the same time, complementing existing imaging resources within the Faculty.

Versatile imaging:  The Nikon A1R offers a unique approach to confocal imaging.  The system incorporates into the scanhead both resonant (fast scanning) and non-resonant (conventional scanning) scanning mirrors and in doing so, allows experimenters to have the option of imaging in high resolution (up to 4096x4096 pixels), at high speed (420fps at 512x32 pixels) or a combination of both.   This arrangement is ideal for live-cell photo-activation and photo-bleaching studies where resonant scanning can be used to capture images at high-speed and non-resonant scanning can be used to target a region of interest for photo-activation / ablation.

LIVE-cell imaging:  Resonant scanning mode has added benefits; for live-cell experiments, photo-sensitive specimens remain live for longer and fluorophore bleaching is significantly reduced.  Of course, the system will also perform regular 2D, 3D and 4D imaging with ease.
Environmentally friendly:  Both A1Rs will incorporate fully-enclosed heated environmental chambers and will have the ability to set, maintain and monitor temperature, humidity, CO2 and O2 levels as imaging conditions dictate.

Spectral imaging:  The A1R comes equipped with spectral detectors to allow the spectral profiling and if required, unmixing of fluorescently ‘complex’ specimens.

Now available: The Nikon A1R is an extremely versatile imaging platform capable of multi-dimensional imaging of cells at both high resolution and speed.  One machine is located in the main Bio-Imaging Unit at the Medical School and the other within the Imaging Suite at the Institute of Genetic Medicine.  Both microscopes are now installed.

TIRFific imaging is now available!

Our spinning disk confocal microscope has undergone some enhancements.  The heart of the system, the Yokogowa CSU10 spinning disk unit, forms the basis of a combination spinning disk confocal and TIRF (Total Internal Reflectance Fluorescence) microscopy setup.  Like the A1R, the tandem system is fully enclosed in an environmental chamber and offers the ability to set humidity, temperature O2 and CO2 levels as required.  The chamber is also blackened so as to allow low light level imaging should the need arise.

TIRF microscopy although not a new imaging technique, has until now, not been unavailable in Newcastle.   TIRF microscopy in combination with the appropriate fluorophore allows the visualization of events taking place in an area within a couple of hundred nanometers of the media-cover slip interface and as such, is an ideal technique with which to visualize membrane –bound events, cell adhesions endo- and exo-cytosis.

The system can be easily switched between TIRF and confocal imaging modes.  It can therefore be used as a versatile live-cell imaging platform and like the A1R, is capable of multi-dimensional high-speed imaging and of course, TIRF.

Nikon BioStation

The Nikon BioStation, a fluorescent imaging platform enclosed within a conventional tissue culture incubator, has recently been upgraded.  As well as a few minor tweaks, the upgrades have improved the BioStation’s ability to perform fluorescence imaging.  Now upgraded, the system is capable of multi-dimensional imaging (2D, 3D, 4D) in phase and fluorescence imaging modes (up to 40x magnification).  The system enables users to image cells over protracted time periods (days-weeks) when time resolution is less important but sample viability is.

With all the above happenings, we hope to be able to offer a series of introductory events, workshops and presentations relating to the new equipment and associated imaging techniques.  Please contact us if you wish to be notified of these events or any bio-imaging related news.

Alex Laude