4/5D Microscopy
The Multi-Dimensional Cell
The capability to perform rapid, multi-dimensional imaging of living cells or model
organisms is becoming increasingly necessary in many areas of modern biological
research. Much of fundamental biological phenomena (spanning development, repair,
cancer, cell cycle, signalling, motility, intracellular transport and viral pathways),
involve microscopic structures moving rapidly through 3D spatial dimensions.
In order to capture these intricate, interactive processes effectively in "real
time", we are required to perform rapid time-lapse imaging of the cell through all
three spatial dimensions i.e. effectively 4D imaging.
Furthermore, since we commonly want to follow the motion and interactions of multiple
intracellular structures, this often requires us to either simultaneously or very
rapidly switch between, the excitation and imaging of several spectrally separated
fluorophore labels i.e. 5D!
Underlying all direct imaging studies of living cells or organisms, is the desire
to preserve the live subject for as long as possible, through minimization of both
phototoxic cell/tissue damage and photobleaching of the incorporated fluorophores.
Techniques that may be employed for 5D cell imaging include spinning disk confocal
microscopy, deconvolution widefield and structured illumination, all of which can
benefit from using Electron Multiplying CCD (EMCCD) technology.
The Multi-Dimensional EMCCD
EMCCD technology is the ideal detector for incorporation into your ultrasensitive
4/D microscopy set-up, whether as a key component in confocal live cell imaging
systems, or as a "stand-alone" EMCCD imaging solution. The extraordinary signal-to-noise
(S/N) ratio offered is significantly greater than that afforded by conventional
point scan detectors. Furthermore, since it is a rapid-readout imaging device the
EMCCD yields rapid frame rates, ideally suited to rapid acquisition of z-stacks
and time-lapse sequences.
The spectral dimension is accessed either through fast switching between multiple
fluorophores, or even splitting of the emission signal simultaneously onto different
areas of the sensor. There is also the possibility to perform true "spectral imaging"
covered elsewhere on this website. Through minimization of excitation powers, the
rates of dye photobleaching and cell phototoxicity are significantly reduced.
Find out more...