Paterson Institute for Cancer Research SCIENTIFIC REPORT 2005

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HEAD OF RESEARCH SERVICES
Jenny Varley
Research Services
http://www.paterson.man.ac.uk/facilities/scifacs.jsp
The Scientific Research Services continue to underpin all the
research activities within the Paterson Institute as well as
providing one CR-UK wide service (the Affymetrix
Microarray service). It is essential that the services provide
comprehensive state-of-the-art facilities which dovetail with
the requirements of the users, and to this end each service
has an associated user group which meets at regular intervals.
These user groups carry significant weight, and can
advise on the introduction of new services or cessation of
redundant ones as well as equipment needs. We have continued
to invest to ensure that all our services are optimally
supported.
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Advanced Imaging Facility
Head: Steve Bagley
http://www.paterson.man.ac.uk/facilities/advimg.jsp
Since 1985 Laser Scanning Confocal Microscopy
has facilitated the digitisation of data in three
dimensions. This has been achieved by the use of a
high intensity monochromatic light source (generally
a laser) and an imaging aperture placed just
before the detector leading to the imaging of those
structures that were in-focus and excluding data
that were above or below the plane of focus. The
system was originally developed to assist imaging at
depths inside embryos that normal microscopy
could not resolve because of interference from
information above and below the plane of focus,
however, the main advance for the broader biological
research community was that 3D imaging
became a routine process available to most laboratories.
Over the last ten years there have been
major advances in optical design, detection sensitivity
and labelling technology which have allowed live
cell imaging via widefield optics to become a viable
possibility.
Widefield imaging requires less light to form an
image hence it is less damaging to cellular processes.
In the last four years the Advanced Imaging
DEPUTY
Caroline Chadwick
Facility has been investigating, designing and developing
techniques for the visualisation of live cellular
processes on the micro- and macro-scale where
the very act of describing real world, four dimensional
objects in a digital manner disrupts biological
processes as little as possible. This has led to a
move away from laser scanning systems to less
harmful and more sensitive techniques utilising
standard optics and image processing algorithms.
Time-lapse of PC3-GFP cells interacting with the BMEC-1 cell layer
over 30 minute time increments.The image shows both the invading
PC3-GFP cells (green) pushing through the mono-layer of
Bodipy Ceramide labelled BMEC-1. Magnification x400
Over the last year we have been increasing the sensitivity
of the available equipment as a matter of
priority. The facility recently purchased a bespoke
multi-position four dimensional microscope, whilst
another has been developed by assembly of optimal
component parts for long term time-lapse; both
allow the collection of several fields of view over
time. This permits questions to be addressed at levels
of temporal and spatial control that were not
possible with previous systems. A considerable
amount of research and development pushed the
system towards two primary goals; 1) to increase the
‘imaging window’ - the available time when biologically
relevant data can be collected whilst reducing
photo-toxicity and 2) to push the sensitivity of the
equipment to image proteins which are present in
low concentration
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