Microplate
assays, e.g. reporter gene assay.
TOP
Cool Imager™
Workstation is a sophisticated device with an innovative
hardware design. See the description below to learn the
details of how the instrument works.
Two-Stage
Pelitier Cooling System
In order to detect low light samples (such as
chemiluminescence) the detection system needs to be very
sensitive. Thermal excitation within the CCD chip, which
converts the light into electric charges, adds electrons to
the detected signal. These extra electrons reduce the
sensitivity. Using a
double-stage
Peltier cooling system, the CCD chip of the Cool
Imager™
Workstation is cooled to -45oC
to avoid ‘thermal noise’ and
achieve maximum sensitivity. In order to achieve such a low
temperature, a vacuum is applied to the camera.
CCD Chip
The central part of the camera system integrated in the
Cool Imager™
is a CCD chip (CCD = Charge-Coupled Device) that converts
incoming light into an electrical signal. The CCD chip is a
small piece of silicon 20.5 mm wide and 16.4 mm high, divided
into 756 rows, each consisting of 581 individual elements,
referred to here as pixels. Each of the pixels is an
individual light detector that can measure the amount of
light hitting the pixel area over a certain period of time.
The number and sensitivity of the pixels define the quality of a
CCD camera. The number of pixels determines the spatial
resolution of the final image. This allows the exact measurement
of very closely spaced bands as well as the use of high density
MTPs.
TOP
The sensitivity of the pixels and the electronics of the
camera determine the lower detection limit and the linearity
of the instrument. During the readout process,
the electric charges collected by the
pixels during exposure are converted into an electrical signal
with 16-bit resolution. The dynamic range of the Cool Imager™
is very wide at 1:10,000. This means
that the Cool Imager™
can measure a 1 pg band on the same
blot as a 10 ng band. By comparison, X-ray film has a low
dynamic range of 1:100. On a blot a 1 pg band and a 100 pg
band could be detected simultaneously, but > 100 pg would not
be distinguishable from 100 pg. Higher concentrations are
often seen as larger bands, though linearity is lost.
This wide dynamic range greatly enhances the
Cool Imager™’s
quantifying ability. In addition, only one exposure is
required to capture faint and strong signals whereas with
X-ray film multiple exposures are required. When data points
are
plotted on a graph and the points are in a straight line, the
data is linear. Therefore linearity refers to how well data
points fall into a straight line. Although linearity is
distinct from dynamic range, the dynamic range of the medium
affects linearity. For example, X-ray film has a poor dynamic
range at 1:100. Even within that dynamic range, the response
of the X-ray film to light, i.e. oxidation of silver grains,
is nonlinear. Thus, X-ray film is not suitable for
quantification. In contrast, the
Cool Imager™
has a dynamic range of 1:10,000 and is linear over this entire
range.
In standard chips,
very strong signals can cause an
electron overflow into neighboring pixels. This can result in
highlighting and flaring of bands. The chip of the Cool Imager™
has a special feature called
antiblooming that allows a pixel to uptake a 1000 x higher
light intensity than a standard chip without overflowing into
neighboring pixels.
TOP
If you want to learn more how a CCD chip works and how the
Cool Imager™
Workstation optimizes the imaging process please continue to
read the next section.
The traditional biological imagers for analyzing DNA gels cannot
be used for the detection of chemiluminescent signals due to
their low sensitivity and high background. In order to detect chemiluminescent signals, the CCD cameras have to be cooled to
a very low temperature to reduce so called "Hot Spots".
Cool Imager™ by Viagene Biotech is integrated with
modern techniques including, computer technology,
microelectronics and semiconductor technology. Cool Imager™
is a digital imager which is used for capturing and analyzing
biologic images with specific design for picking up
chemiluminescent signals. The comparison of it with other
devices on the market is shown in Table 1
and 2.