User manual | pco.edge 4.2

pco.edge 4.2
scientific CMOS camera
low noise
high resolution
0.8 electrons
2048 x 2048 pixel
lightsheet
scanning
mode
USB 3.0
Camera Link
small
form factor
high speed
100 fps
high dynamic range
37 500 :1
high quantum efficiency
up to 82 %
pco.
pco.edge 4.2 | scientific CMOS camera
features
Selectable rolling shutter operation modes of pco.edge cameras.
dual outside in
dual top down
dual inside out
single top down
rolling shutter readout modes - optimized for synchronization of microscopes and scanning applications
All pco.edge sCMOS cameras from the beginning
feature a variety of precise synchronization modes,
which are optimized for advanced microscopy imaging and scanning. The flexible frame and line triggers with very low latency in combination with the
free selectable readout modes can easily be combined to cover every modern microscopy situation
to name a few:
n
n
n
n
n
n
For example, one mode is used in a lightsheet or
SPIM application, the lower right rolling shutter
operational mode “single top down” operation is
convenient to properly synchronize the camera
exposure with the scanner. On the other hand,
if speed is required and a flash like exposure is
applied the upper left mode “dual outside in” is
used for localization microscopy techniques like
GSD, PALM or STORM.
lightsheet microscopy
selective plane imaging microscopy (SPIM)
structured illumination microscopy
localizations microscopy
(GSD, PALM, STORM, dSTORM)
spinning disk confocal microscopy
RESOLFT
pco.
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pco.edge 4.2 | scientific CMOS camera
features
free of drift
The pco.edge sCMOS cameras feature temperature stabilized Peltier cooling, allowing for continuous operation free of drift phenomena in image
sequences capture. This is achieved by the proper
selection and sophisticated combination of electronics and FPGA algorithms.
As the measurement result shows while running at
full speed of 100 frames/s over 4 hours measuring
time the camera doesn’t show any significant drift
(figure on the right side). This degree of stability
enables long-term measuring series, which should
be quantitatively evaluated and processed. For
example, in PCR (Polymerase Chain Reaction)
applications, when so-called melting curves must
be measured, the fluorescence in multi-well plates
with different samples is recorded over a longer
time at different sample temperatures. Here all
the images are used for processing, which is only
possible if the offset is stable and the camera is
free of drift.
Mean dark signal drift measurement of a pco.edge camera stabilized
at +5 °C over a 4 hour period recorded at 100 frames/s
(1 count = 0.5 electron).
reaching emCCD domain
The graph shows the signal-to-noise (SNR) curves of a typical
emCCD camera (gain = 1000) and a pco.edge 4.2 camera vs.
number of photons.
pco.
In the past emCCD image sensors featuring on-chip
amplification were developed to detect the lowest level of light. However, amplification, while reducing read
out noise, comes at the expense of dynamic range.
Both features are not possible simultaneously in
emCCD sensors. In addition, the amplification process
generates excess noise, which reduces the effective
quantum efficiency (QEeff) of the emCCD sensor by
the factor of two (e.g. the 90 % QE of a back illuminated emCCD sensor has an QEeff of 45 %). The excess
noise present in emCCDs makes the pco.sCMOS the
sensor of choice at light conditions above 1 photon
per pixel (at 80 % QE, assuming a cooled sensor with
dark current = 0). Furthermore, available emCCD sensors are limited in resolution and frame rate.
3
pco.edge 4.2 | scientific CMOS camera
features
readout noise in sCMOS
The EMVA 1288 standard explains that in principle
for each pixel in an image sensor the noise behavior
is determined by recording many images and calculating the time dependent variation or deviation of
each pixel from its mean value. This is the determination of the root mean square (rms) value for each
pixel. Since the widely used CCD image sensors
don’t have a separate output stage for each pixel,
the variation of the noise between each pixel is minimal. Therefore, instead of measuring many images,
it is sufficient to measure two images, calculate the
variance for each pixel and average these variances
within the image to obtain an rms value for the image sensor. For CCD image sensors this simplification is a good approximation and has been now for
years to describe the readout noise of image sensors in general.
However, CMOS image sensors, including scientific
CMOS image sensors, feature a different behavior
such that the simplified rms determination with the
averaging across the whole image sensor is not sufficient to describe the noise behavior. The figure top
right shows the result of time series of dark images,
where for each pixel an rms value is calculated along
the time axis and the results are shown in this histogram, showing the readout noise distribution for the
total image sensor. Since two different pixel clocks
are available in turn two curves are provided.
Noise distribution of the rms raw data values (noise filter off) of each
pixel in the dark image of a pco.edge 4.2 at different readout speeds
(slow scan / fast scan).
A valuable characterization of these rms value distributions is the so called median value, which is the
point where 50% of all values are larger and smaller.
For comparison the rms value measured by the simplified EMVA1288 approach is given. For a CCD image sensor these values would be identical, but for
CMOS image sensors they start to diverge. For comparison of different cameras and image sensors both
values can be used. For practical use it should be
considered, that these values are calculated from a
large series of recorded images.
The left figure shows the same fast scan curve of the
pco.edge 4.2 only in a logarithmic y-axis (frequency)
scaling, to emphasize that most of the pixels have an
average readout noise in time that is smaller than 1
electron and there are few pixels (less than 1 % of
the maximum), which have a readout noise of 3 – 6
electrons.
Noise distribution of the rms raw data values (noise filter off) of
each pixel in the dark image of a pco.edge 4.2 at the fast readout
speed. Graph is identical to figure on the top but in logarithmic
y- axis scaling.
pco.
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pco.edge 4.2 | scientific CMOS camera
features
superior image quality
The pco.edge sCMOS camera features outstanding low read out noise. Even at maximum speed of
100 frames/s at full resolution of 2048 x 2048 pixel
the noise is 0.9 e- med. Moreover the pco.edge provides an excellent homogeneous pixel response to
light (PRNU, photo response non-uniformity) and an
excellent homogeneous dark signal pixel behaviour
(DSNU, dark signal non-uniformity), which is achieved
by a sophisticated electronic circuit technology and
firmware algorithms.
The lower figure shows a comparison of a scientific
grade CCD and the new pco.sCMOS image sensor
under similar weak illumination conditions. This demonstrates the superiority of sCMOS over CCD with
regards to read out noise and dynamic, without any
smear (the vertical lines in the CCD image).
Dark image comparison with the measured distribution of “hot
blinking” pixels at 5°C of the image sensor. The left image gives a
3D view with the sophisticated “blinker filter” algorithm off and the
right image shows the result with the filter switched on.
The left image was recorded by a scientific CCD camera while the
right image was recorded by a pco.edge under identical conditions.
flexibility and free of latency
User selectable choice of rolling shutter modes for exposure provides flexibility for a wide range of applications.
The advantages of rolling shutter are high frame rates and low read out noise. In Camera Link Version due to
realtime transmission of the image data to the PC, there is no latency between recording and access or storage
of the data.
37 500:1 dynamic range
The top image shows an extract of a typical pco.edge recording of
a grey scale with a 1 : 10 000 dynamic in 20 steps.
The bottom image is a plot of the grey values profile along the
centered line through the top image (with gamma 2.2).
pco.
Due to the excellent low noise and the high fullwell
capacity of the sCMOS image sensor an intra scene
dynamic range of better than 37 500 : 1 is achieved
(with 100 fps and fast scan read out mode). A unique
architecture of dual column level amplifiers and dual
11 bit ADCs is designed to maximize dynamic range
and to minimize read out noise simultaneously. Both
ADC values are analyzed and merged into one high
dynamic 16 bit value.
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pco.edge 4.2 | scientific CMOS camera
features
high resolution
A 4.2 Mpixel resolution in combination with a moderate chip size (18.8 mm diagonal, 6.5 μm pixel pitch)
benefits microscopy applications with low magnification factor and large field of view, thereby reducing
processing times and increasing throughput. The
figure compares the potential of the new field of view
of the pco.edge to the 1.3 Mpixel image resolution
which is widely used in microscopy applications for
scientific cameras.
The two images show in comparison the field of view with sCMOS
resolution vs. a 1.3 Mpixel resolution, courtesy of Dr. Stefan Jakobs,
Dept. of NanoBiophotonics, MPI for Biophysical Chemistry
high speed recording and data streaming
The new pco.edge offers in fast mode a frame rate of 100 frames/s (fps) at full resolution of 2048 x 2048 pixel
as a full download stream to the PC. Therefore the recording time is just limited by either the amount of RAM
in the PC or, in case of a RAID system, by the capacity and number of hard disks. As in many CMOS based
cameras the frame rate increases significantly if smaller regions of interest (ROI) are used. The reduction of the
image area works as well in favour of the frame rate of CCD sensors, but here unwanted regions still need to be
read out at the expense of the total readout speed. The typical frame rate for a 1.3 Mpixel scientific CCD camera
(6 e- read out noise) is 10 fps. The new pco.edge camera provides at 1.3 Mpixel resolution (< 1.0 e- readout
noise) a frame rate of 200 fps in comparison.
pco.
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pco.edge 4.2 | scientific CMOS camera
the new interface standard
for ultimate performance cameras
What is Camera Link HS?
Camera Link HS (CLHS) is designed to specifically meet the needs of vision and imaging
applications. It provides low latency, low jitter, real-time signals between a camera and a
frame grabber while carrying image data, control data and trigger events. The interface
builds upon the key strengths of Camera Link by adding new features and functions
to meet the needs of today and tomorrow. Camera Link HS is designed as a system
ensuring that CMOS sensor technology can be fully exploited, while providing
cost effective cameras and frame grabbers that are easy to use, flexibility and
provide reliability data demanded by customers1.
Benefits
1
http://www.visiononline.org/vision-
standards-details.cfm?type=10
More bandwidth
• Effective bandwidth of about 1187 MB/s (CLHS X-Protocol - 10G) equals roughly
three times a USB 3.1 Gen1 bandwidth & equals netto data rate of CoaXPress CXP-12
More robust connection
• No communication error at a Bit Error Rate (BER) of 10-12
because of using a Forward Error Correction algorithm (FEC)
• Forward Error Correction corrects burst errors of up to 11 Bits on the fly
• FEC technology supersedes packet resend mechanism for data reliability
• Fiber Optic Link (FOL) provides high resistance to EMC and allows
long cable lengths with the best signal integrity
More distance
• Cable length more than 300m using multimode fiber
• Cable length more than 10km with single mode fiber
More flexibility
• Real-time trigger over cable with extremely low jitter
• Plug and Play with GenICam and GenCP
• Using standard LC-connector for flexible cable decision
More open
• The full CLHS specification is downloadable for free
• AIA IP-core is available for fast compliant FPGA implementation (Xilinx, Altera, Lattice)
More cost effective
• The use of standard network hardware components such as enhanced small
form-factor pluggable (SFP+) connector from multiple vendors allows multi sourcing
• Inexpensive licensing
1 http://www.visiononline.org/vision-standards-details.cfm?type=10
pco.
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pco.edge 4.2 | scientific CMOS camera
technical data
Camera Link HS
camera
image sensor
type of sensor
image sensor
resolution (h x v)
pixel size (h x v)
sensor format / diagonal
shutter modes
MTF
fullwell capacity (typ.)
readout noise1
dynamic range (typ.)
quantum efficiency
spectral range
dark current (typ.)
DSNU
PRNU
anti blooming factor
scientific CMOS (sCMOS)
CIS2020A
2048 x 2048 active pixel
6.5 µm x 6.5 µm
13.3 mm x 13.3 mm / 18.8 mm
rolling shutter (RS)
with free selectable readout modes
76.9 lp/mm (theoretical)
30 000 e0.8med /1.3rms e- @ slow scan
0.9med /1.4rms e- @ fast scan
37 500 : 1 (91.5 dB) slow scan
up to 82 % @ peak
370 nm .. 1100 nm
< 0.6 e-/pixel/s @ 7 °C
< 0.3 e- rms
< 0.3 %
> 10 000
frame rate
@ 2048 x 2048 pixel
exposure / shutter time
dynamic range A/D2
A/D conversion factor
pixel scan rate
pixel data rate
binning horizontal
binning vertical
region of interest (ROI)
non linearity
cooling method
trigger input signals
trigger output signals
frame rate table3
typical examples
fast scan
slow scan
2048 x 2048
2048 x 1024
2048 x 512
2048 x 256
2048 x 128
100 fps
200 fps
400 fps
800 fps
1600 fps
35 fps
70 fps
140 fps
281 fps
562 fps
1920 x 1080
1600 x 1200
1280 x 1024
640 x 480
320 x 240
189 fps
170 fps
200 fps
420 fps
853 fps
66 fps
60 fps
70 fps
150 fps
300 fps
data interface
time stamp
100 fps @ RS, fast scan
100 µs .. 10 s (RS)
16 bit
0.46 e-/count
274.0 MHz fast scan
100.0 MHz slow scan
548.0 Mpixel/s
200.0 Mpixel/s
x1, x2, x4
x1, x2, x4
horizontal: steps of 16 pixels
vertical: steps of 1 pixel
< 0.5 %
+ 7°C stabilized
peltier with forced air (fan)
(up to 27°C ambient)
frame trigger, sequence trigger,
programmable input (SMA connectors)
exposure, busy, line, programmable
output (SMA connectors)
Camera Link HS (Single-F2,1X1,S10)
in image (1 µs resolution)
general
power supply
power consumption
weight
operating temperature
operating humidity range
storage temperature range
optical interface
CE / FCC certified
12 .. 24 VDC (+/- 10 %)
32 W max. (typ. 19 W @ 20 °C)
1010 g with F-mount
+ 10 °C .. + 40 °C
10 % .. 80 % (non-condensing)
- 10 °C .. + 60 °C
F-mount & C-mount
yes
1 The readout noise values are given as median (med) and root mean square (rms) values, due to the
different noise models, which can be used for evaluation. All values are raw data without any filtering.
2 The high dynamic signal is simultaneously converted at high and low gain by two 11 bit A/D converters
and the two 11 bit values are sophistically merged into one 16 bit value.
3 Max. fps with centered ROI.
pco.
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pco.edge 4.2 | scientific CMOS camera
technical data
Camera Link
camera
image sensor
type of sensor
image sensor
resolution (h x v)
pixel size (h x v)
sensor format / diagonal
shutter modes
MTF
fullwell capacity (typ.)
readout noise2
dynamic range (typ.)
quantum efficiency
spectral range
dark current (typ.)
DSNU
PRNU
anti blooming factor
scientific CMOS (sCMOS)
CIS2020A
2048 x 2048 active pixel
6.5 µm x 6.5 µm
13.3 mm x 13.3 mm / 18.8 mm
rolling shutter (RS)
with free selectable readout modes,
lightsheet scanning mode1
76.9 lp/mm (theoretical)
30 000 e0.9med /1.4rms e- @ slow scan
1.0med /1.5rms e- @ fast scan
33 000 : 1 (90.4 dB) slow scan
> 82 % @ peak
370 nm .. 1100 nm
< 0.5 e-/pixel/s @ 5 °C
< 1.0 e- rms
frame rate
@ 2048 x 2048 pixel
exposure / shutter time
dynamic range A/D5
A/D conversion factor
pixel scan rate
pixel data rate
binning horizontal
binning vertical
region of interest (ROI)
non linearity
cooling method
< 0.5 %
> 10 000
trigger input signals
trigger output signals
frame rate table3
typical examples
fast scan
slow scan
2048 x 2048
2048 x 1024
2048 x 512
2048 x 256
2048 x 128
100 fps
200 fps
400 fps
800 fps
1600 fps
35 fps
70 fps
140 fps
281 fps
562 fps
1920 x 1080
1600 x 1200
1280 x 1024
640 x 480
320 x 240
189 fps
170 fps
200 fps
420 fps
853 fps
66 fps
60 fps
70 fps
150 fps
300 fps
data interface
time stamp
100 fps, fast scan
100 µs .. 10 s
16 bit
0.46 e-/count
272.3 MHz fast scan
95.3 MHz slow scan
544.6 Mpixel/s
190.7 Mpixel/s
x1, x2, x4
x1, x2, x4
horizontal: steps of 1 pixel
vertical: steps of 1 pixel
<1%
+ 5 °C stabilized
selectable:
peltier with forced air (fan)
or water cooling
(both up to 27°C ambient)
frame trigger, sequence trigger,
programmable input (SMA connectors)
exposure, busy, line, programmable
output (SMA connectors)
Camera Link Full (10 taps, 85 MHz)
in image (1 µs resolution)
general
power supply
power consumption
weight
operating temperature
operating humidity range
storage temperature range
optical interface
CE / FCC certified
12 .. 24 VDC (+/- 10 %)
20 W max. (typ. 10 W @ 20 °C)
700 g
+ 10 °C .. + 40 °C
10 % .. 80 % (non-condensing)
- 10 °C .. + 60 °C
F-mount & C-mount
yes
frame rate table extended readout mode4
typical examples
fast scan
slow scan
2048 + 12 x 2048
2048 + 12 x 1024
100 fps
200 fps
35 fps
70 fps
1 Selectable via SDK (software development kit).
2 The readout noise values are given as median (med) and root mean square (rms) values, due to the
different noise models, which can be used for evaluation. All values are raw data without any filtering.
3 Max. fps with centered ROI.
lightsheet
scanning
mode
4 Extended readout mode with 12 columns of black reference pixel.
5 The high dynamic signal is simultaneously converted at high and low gain by two 11 bit A/D converters
and the two 11 bit values are sophistically merged into one 16 bit value.
pco.
9
pco.edge 4.2 | scientific CMOS camera
technical data USB 3.0
camera
image sensor
type of sensor
scientific CMOS (sCMOS)
image sensor
resolution (h x v)
pixel size (h x v)
sensor format / diagonal
shutter modes
CIS2020A
2048 x 2048 active pixel
6.5 µm x 6.5 µm
13.3 mm x 13.3 mm / 18.8 mm
rolling shutter (RS)
with free selectable readout modes,
global reset - rolling readout (GR)
76.9 lp/mm (theoretical)
30 000 e0.8med /1.3rms e37 500 : 1 (91.5 dB)
up to 82 % @ peak
370 nm .. 1100 nm
< 0.3 e-/pixel/s @ 0 °C
< 0.3 e- rms
MTF
fullwell capacity (typ.)
readout noise1
dynamic range (typ.)
quantum efficiency
spectral range
dark current (typ.)
DSNU
PRNU
anti blooming factor
< 0.2 %
> 10 000
frame rate
@ 2048 x 2048 pixel
exposure / shutter time
dynamic range A/D3
A/D conversion factor
pixel scan rate
pixel data rate
binning horizontal
binning vertical
region of interest (ROI)
non-linearity
cooling method
trigger input signals
trigger output signals
data interface
time stamp
frame rate table2
100 µs .. 20 s RS
30 µs .. 2 s GR
16 bit
0.46 e-/count
110.0 MHz
220.0 Mpixel/s
x1, x2, x4
x1, x2, x4
horizontal: steps of 4 pixels
vertical: steps of 1 pixel
< 0.6 %
0 °C stabilized, peltier with
forced air (fan) / water cooling
(both up to 27°C ambient)
frame trigger, programmable input
(SMA connectors)
exposure, busy, line, programmable
output (SMA connectors)
USB 3.0
in image (1 µs resolution)
general
typical examples
2048 x 2048
2048 x 1024
2048 x 512
2048 x 256
2048 x 128
40 fps
80 fps
160 fps
315 fps
610 fps
1920 x 1080
1600 x 1200
1280 x 1024
640 x 480
320 x 240
76 fps
69 fps
80 fps
170 fps
335 fps
pco.
40 fps
power supply
power consumption
weight
operating temperature
operating humidity range
storage temperature range
optical interface
CE / FCC certified
12 .. 24 VDC (+/- 10 %)
21 W max. (typ. 12 W @ 20 °C)
930 g
+ 10 °C .. + 40 °C
10 % .. 80 % (non-condensing)
- 10 °C .. + 60 °C
F-mount & C-mount
yes
1 The readout noise values are given as median (med) and root mean square (rms) values, due to the
different noise models, which can be used for evaluation. All values are raw data without any filtering.
2 Max. fps with centered ROI.
3 The high dynamic signal is simultaneously converted at high and low gain by two 11 bit A/D converters
and the two 11 bit values are sophistically merged into one 16 bit value.
10
pco.edge 4.2 | scientific CMOS camera
technical data
quantum efficiency
monochrome
camera views
Camera Link HS
Camera Link
USB 3.0
dimensions
F-mount and C-mount lens changeable adapter.
~31 (adjustable)
3x 1/4" 20 UNC
70
Rubber feet
C-Mount
38,20
3,50
F-Mount
48
76
pco.edge
Camera Link HS
~1,8 (adjustable)
122,50
88,90
122,50
pco.
3x 1/4" 3x
3x201/4"
20 1/4"
20 UNC
UNC
UNC
70
48
48
48
76
C-Mount
C-Mount
C-Mount
3,50
38,20 38,20 38,20
3,50
All dimensions are given in millimeter.
70
RubberRubber
feet Rubber
feet feet
76
76
F-MountF-Mount
F-Mount
3,50
pco.edge
Camera Link/
USB 3.0
70
~1,8(adjustable)
~1,8(adjustable)
~1,8(adjustable)
~31(adjustable)
~31(adjustable)
~31(adjustable)
99,50 99,50 99,50
88,90 88,90 88,90
99,50 99,50 99,50
11
pco.edge 4.2 | scientific CMOS camera
technical data
software
third party integrations
software drivers
For camera control, image acquisition and archiving
of images in various file formats PCO provides the
software application Camware (Windows 7, 8 and later).
A camera SDK (software development kit) including a
32 / 64 bit dynamic link library for user customization
and integration on PC platforms is available for free.
For camera interface drivers and a list of supported
third party software please visit www.pco.de.
options
custom made versions (e.g. water cooling)
Water cooling unit Aquamatic II
for use with pco.edge cameras.
pco.
12
pco.edge 4.2 | scientific CMOS camera
applications
life science
physical science
life science
A widefield (right) and a GSDIM superresolution (left) microscopy image of tubulin
fibers obtained with a pco.edge, courtesy
of Leica Microsystems, Germany
A single image of fluorescence labeled
protein networks in water drops in an
oil phase, which moved fast. One pixel
corresponds to 0.1625 µm in reality,
courtesy of Prof. Dr. Sarah Köster, Institute
for X-Ray Physics, Göttingen, Germany
Zebrafish with two fluorescent labels,
collected with a VisiScope Confocal based
on the Yokogawa CSU-W1 wide head and
a pco.edge camera, courtesy of Visitron
Systems GmbH, Germany
life science
life science
life science
Neuronal network marked with a
fluorophore (false color rendering) and
recorded with a pco.edge
Extract of a fluorescent slide which was
scanned by a pco.edge camera in a
Pannoramic 250 Flash scanner for digital
pathology, courtesy of 3DHistech, Hungary
An image of a sequence, which was
recorded with a pco.edge at 400 frame/s.
The maximum signal was about 100
photons, courtesy of Prof. Engstler,
University of Würzburg, Germany
application areas
n Widefield microscopy n Fluorescent microscopy n Digital pathology n PALM n STORM n GSDIM
n dSTORM n Superresolution microscopy n Lightsheet microscopy n Selective plane imaging microscopy
(SPIM) n Calcium imaging n FRET n FRAP n 3D structured illumination microscopy n High speed bright
field ratio imaging n High throughput screening n High content screening n Biochip reading n TIRF n TIRF
microscopy / waveguides n Spinning disk confocal microscopy n Live cell microscopy n 3D metrology n
TV / broadcasting n Ophtalmology n Electro physiology n Lucky astronomy n Photovoltaic inspection
europe
america
asia
PCO AG
Donaupark 11
93309 Kelheim, Germany
PCO-TECH Inc.
6930 Metroplex Drive
Romulus, Michigan 48174, USA
PCO Imaging Asia Pte.
3 Temasek Ave
Centennial Tower, Level 34
Singapore, 039190
fon +49 (0)9441 2005 50
fax +49 (0)9441 2005 20
[email protected]
www.pco.de
fon +1 (248) 276 8820
fax +1 (248) 276 8825
[email protected]
www.pco-tech.com
fon +65-6549-7054
fax +65-6549-7001
[email protected]
www.pco-imaging.de
pco.
subject to changes without prior notice | ©PCO AG, Kelheim | pco.edge 4.2 | v1.02A
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