The iXon3 is capable of market leading frame rate performance, achieved from ‘over-
clocking’ the vertical shifts during readout. Furthermore, fastest possible continuous sub-
region frame rates can be attained using the new ‘cropped sensor’ mode.
Since the launch in late 2010 of imaging cameras that are based on a new 5.5 megapixel scientific CMOS
(sCMOS) sensor, there has been much speculation about whether or not sCMOS will be seen as a technology
replacement for interline CCD and electron multiplying CCD (EMCCD) cameras – which, in many ways,
can be considered the current gold standards for low light fluorescence microscopy and bio-imaging in
general. Coming from the unique market position of manufacturing all of the aforementioned camera types,
we provide here an analysis of how these sensitive imaging technologies compare.
OptAcquire is a unique control interface, whereby a user can conveniently choose from
a pre-determined list of set-up configurations, each designed to optimize the camera for
different experimental acquisition types, thus removing complexity from the extremely
adaptable control architecture of the iXon3.
The organization of a cell is critical for its function and understanding how organization
affects function is a major goal of cell biology. Researchers led by Dr. John A. Cooper at
the University of Washington in St. Louis and Dr Alexey Khodjakov at the Wadsworth
Center, Albany, New York used microtubule ablation to learn more about how cells use the
cytoskeleton to integrate spatial information into cell cycle regulation.
Scientists can learn about the functions of specific neurons in networks, like the
brain using proteins that bind natural or synthetic photoswitches. Light can turn these
photoswitches on and off, allowing the control of activity in specific cells and thus
observation of how processing and behavior are altered by defined neuronal populations.
Much has been learned about cells and their proteins using optical reporters such as GFP,
but scientists want to know more. Work is now aimed at using light not only to observe
cellular functions but also to control them in very precise ways. Optical manipulation of
cell function is being exploited in cutting edge research to control behavior in organisms,
characterize signaling pathways and test cellular network models. The approach has been
dubbed “Optogenetics” and was named “Method of the Year 2010” by Nature publishing.
Calcium ions are known to play a role in many aspects of cell function, but gaps exist
in the understanding of calcium’s role in the cell cycle. Research led by Professor Mike
White and Dr. Violaine Sée at the University of Liverpool used targeted flash photolysis
to elucidate intracellular calcium’s role in signaling the cell to progress through the cell
cycle.
Intracellular pH is important in many functions that take place in a cell. For example, pH
affects protein structure and the function of lysosomes, mitochondria, and other organelles.
Changes in energy metabolism also often correlate with pH changes, and so scientists
would like to monitor ATP and pH in a cell simultaneously.
Prof. Dr. Ben L. Feringa and Dr. Wesley R. Browne from the University of Groningen,
the Netherlands, are using synthetic chemistry to create new light-responsive nanoscale
structures that could one day find use in applications such as smart materials and drug
delivery. The photoreactivity of the structures being developed places the Andor Revolution
DSD confocal microscope as a key asset in studying the dynamic properties of the materials
in real time.
One of the distinctive features of the iXon3 is the capability to quantitatively capture and
present data in units of electrons or photons, the conversion applied either in real time or
as a post-conversion step.
Calculating dynamic range in EMCCDs has often been a source of confusion, due to the
additional requirement to factor in EM gain and the extended well capacity of the gain
register. High dynamic range can be accessed in EMCCDs with careful fine tuning
of EM gain.
Fast Kinetics Acquisition Mode can be used to acquire bursts of data with sub-
microsecond time resolution. The iXon3 is configured to make available not only the rows
of the image area, but also rows under the frame transfer mask for storage of acquired data
prior to readout. The ‘overclocked’ vertical shift speeds of the iXon3 renders it ideal for
extremely fast temporal resolution in Fast Kinetics Mode.
A true scientific CMOS in every sense, Neo has been conceptualised and specifically engineered to harness the full performance potential of this new and exciting sensor technology. Unlike any CCD or CMOS camera to come before, Neo is unique in its ability to simultaneously offer ultra-low noise, extremely fast frame rates, wide dynamic range, high resolution and a large field of view.
In this article, we describe some novel camera-based approaches which enable the cell biologist to break through the diffraction-limited spatial resolution of classical light microscopy, enabling much finer sub-cellular detail to be elucidated than was previously possible. The role of ultra-sensitive, rapid readout cameras is proving central to the ability to drive these methods faster, particularly critical to realize the goal of super-resolution imaging of dynamic processes in living cells.
Dr. Colin Coates discusses Andor's EMCCD Cameras for Low Light Microscopy.