ORCA-Flash4.0 V2 C11440-22CUDigital CMOS camera
Two Scan Speeds
While the read noise at standard scan is only 1.9 electrons rms (1.3 electrons median), there are some experiments for which even lower noise is more important than raw speed. New in the ORCA-Flash4.0 V2 is an additional slow scan readout mode with read noise of just 1.5 electrons rms (0.9 electrons median). Both the USB and Camera Link configurations of the camera have this low noise capability.
Lightsheet Readout Mode (Patent pending)
To enable the best speeds and synchronization for light sheet microscopy, the ORCA-Flash4.0 V2 configured with the Camera Link interface can be read out in one sweep across the sensor from top to bottom or bottom to top using our new Lightsheet Readout Mode.
Global Exposure Flexibility
By adding a Global Reset function to the ORCA-Flash4.0 V2, users can acquire global exposures and choose to have either an external source or the camera be master of the timing.
Knowing as much as possible about your camera helps increase confidence in the results it produces - especially under demanding experimental conditions. Every ORCA-Flash4.0 V2 is individually characterized at the factory before it ships, and the results of these tests are included with each camera. A measured noise histogram, photon transfer curve, rms noise value and conversion factor (electron/count) are provided along with simple formulas to make use of this information. Next time you’re asked how many photons were detected you’ll know the answer!
High sensitivity means extreme versatility
The ORCA-Flash4.0 V2 is changing the game of scientific imaging. For years, cooled CCDs have been the go-to technology for fluorescence applications such as GFP or multi-channel imaging that require high signal to noise, high contrast images. EM-CCDs have been scientists' choice for low-light, often high speed applications such as TIRF or spinning disk confocal. For lack of a better choice, the same technology has been adopted for localization microscopy. The ORCA-Flash4.0 V2 offers such a multitude of benefits that it not only easily accomplishes each of these applications -- it may do them better.
|Fan Long, Shaoqun Zeng, and Zhen-Li Huang. "Localization-based super-resolution microscopy with an sCMOS camera Part II:
Experimental methodology for comparing sCMOS with EMCCD cameras," Optics Express, Vol. 20, Issue 16, pp. 17741-17759 (2012)
Quantum efficiency: higher than 70 % at 600 nm and 50 % at 750 nm
The ORCA-Flash4.0 V2 is engineered to outperform all other cameras for fluorescence microscopy. With carefully designed pixels and on-chip lens technology, its Gen II sCMOS sensor provides high QE across the range of wavelengths most commonly used in fluorescence microscopy.
The ORCA-Flash4.0 V2 has the lowest read noise at 100 frames/s of any CCD or sCMOS camera. Even EM-CCDs trade off “relative” low read noise for multiplicative noise by using on-chip gain. But the ORCA-Flash4.0 V2 requires no tradeoffs. Our “quiet” electronics successfully lower the limit of detection, allowing you to take full advantage of high frame rates and see your signal with fewer photons.
The unique combination of high quantum efficiency and low noise, in the absence of EM-CCD multiplicative noise, means that your images are not limited by the camera. Detect signal at low light levels, compare small changes in intensity, and discriminate small signals amid large backgrounds—with ease.
Wide field of view & high resolution
With 4.0 megapixels at 6.5 μm × 6.5 μm each, the ORCA-Flash4.0 V2 is the ideal format for demanding microscopy applications. Whether imaging at high magnification, requiring finely detailed images of an individual cell, or low magnification, aiming to capture and resolve images of many cells, the ORCA-Flash4.0 V2 delivers beautiful images.
High speed Up to 40 min. of continuous full speed, full resolution acquisition*1
Allegro or presto? You be the Conductor.
When conducting imaging with a camera that has 4 194 304 pixels with 16-bit data depth, a single image is 8 megabytes. But capturing a single frame is child's play. What really matters is sustained, sequential image capture. Hamamatsu's ImageConductor gives you control over which speed works for you. In the default configuration, the ORCA-Flash4.0 V2 comes with a USB 3.0 card and cable and will deliver 30 frames/s of full frame acquisition. If you choose, upgrade to our fully supported Firebird PCI Express Gen II 8× Camera Link card, and that very same camera, without any additional modifications, can achieve 100 frames/s full resolution speed. Combining the Camera Link version with our recommended solid state hard drive and high-speed computer keeps your data flowing, for up to 40 minutes of full speed, full resolution recording. Both camera configurations facilitate fine tuning of frame rates by allowing flexible region of interest, letting you select the area that matters. At all speeds, in every configuration, the ORCA-Flash4.0 V2 has just 1.9 electrons rms (1.3 electrons median) read noise for the ultimate in versatility and performance.
*1 This was tested with Dell T5500 (E5640 2.66GHz)+RAID0 (LSI MegaRAID SAS 9260-4i) and 4 pcs SATA SSD drivers (SAMSUNG MZ-7PC512) Windows7 64 bit
Conduct your research
Every ORCA-Flash4.0 V2 includes ImageConductor connectivity™ so that it’s enabled for both USB 3.0 (default) and high speed Camera Link. If your imaging tempo is 30 frames/s, then the default configuration with USB 3.0 is right for you. If you need something a little more lively… presto, just add a Camera Link board now or later to achieve 100 frames/s of full 4-megapixel images. Both options deliver the same low noise, high quantum efficiency imaging for unprecedented sensitivity. With Hamamatsu’s versatile ImageConductor connectivity™ you direct the show.
Read noise: rms or median?
RMS and median are both valid statistical models for evaluating the central tendencies of data distributions, such as pixel noise. With CCDs there are never any issues regarding which model to use because the typical read noise for all pixels is very similar; thus rms and median are equivalent. With sCMOS, the structure of the sensor inherently has more pixel variation and the extreme low noise of the sensor makes variation more statistically significant. But when it comes to evaluating camera performance, the truly meaningful spec is rms noise. The rms noise value provides insight into image quality as well as being the appropriate noise variable in quantitative calculations. The ORCA-Flash4.0 V2's median noise data of 1.3 electrons (typical) is included only to facilitate apparent comparison with other sCMOS cameras. For truly quantitative imaging, rms noise must be known. The ORCA-Flash4.0 V2 Gen II sCMOS has 1.9 electrons rms typical read noise.
All pixels or some pixels?
RMS or median noise values are valid only if all the pixels in the sensor are used or if the exclusion of outlier pixels is documented and explained. For the ORCA-Flash4.0 V2, we calculate both the rms and median read noise using every pixel in the sensor. This is done without any pixel correction functions or prequalification of the data. Since one goal of providing a spec is to enable accurate quantification of imaging results, this approach is consistent with our goal of providing the best quantitative scientific cameras.
Thinking in photons
There is a disconnect in imaging: we image photons but we talk about camera specs in electrons. This gap can be bridged easily, making camera comparisons more meaningful. Consider the difference between Gen I sCMOS and Gen II sCMOS. On the face of it, read noise specs seem rather equivalent (be careful to compare rms to rms, median to median under analogous modes and speeds). But if the read noise is considered first in electrons rms and then converted to photons, using QE at a particular wavelength, the differences are pronounced. At 100 frames/s in rolling shutter mode, the Gen II ORCA-Flash4.0 V2 has 1.9 electrons rms while Gen I has 2 electrons rms. The QEs at 550 nm for Gen I and Gen II are 54 % and 72 %, respectively. Using these numbers, the read noise in photons for Gen I is 2/0.54 = 3.7, while the Gen II is just 1.9/0.72 = 2.6. So now, in photons, the Gen I sCMOS has 42 % higher read noise than the ORCAFlash4.0 V2.
Finally, consider the outcome of this exercise when the ORCA-Flash4.0 V2 is running in slow scan mode with only 1.5 electrons rms noise. The read noise is a mere 2.08 photons.
The ORCA-Flash4.0 V2 is ideally suited for fluorescence and other widefield microscopy applications.
· Super-resolution microscopy
· TIRF microscopy
· Ratio imaging
· High-speed Ca2+ imaging
· Real-time confocal microscopy
· Light sheet microscopy
With the introduction of the ORCA-Flash4.0 V2, users are now able to stream 4 megapixel images to their computers 100 frames per second. The computer requirements for this high data rate can be met by using the guidelines listed this PC Recommendations for ORCA-Flash4.0 V2.
PC Recommendations for ORCA-Flash4.0 V2 [1 MB/PDF］
|Quantum efficency||Higher than 70 % at 600 nm and 50 % at 750 nm|
|Imaging device||Scientific CMOS Sensor FL-400|
|Effective no. of pixels||2048 (H)×2048 (V)|
|Cell size||6.5 μm×6.5 μm|
|Effective area||13.312 mm×13.312 mm|
|Full well capacity||30 000 electrons (typ.)|
|Readout speed||100 frames/s (Full resolution, Camera Link)
30 frames/s (Full resolution, USB 3.0)
|Readout noise||Standard scan (at 100 frames/s, typ.):1.9 electrons rms (1.3 electrons median)
Slow scan (at 30 frames/s, typ.): 1.5 electrons rms (0.9 electrons median)
|Exposure time||Internal trigger mode: 1 ms to 10 s (at full resolution)*1
Internal trigger mode with sub-array readout: 38.96 μs to 10 s
External trigger mode with sub-array readout: 1 ms to 10 s
|Cooling method||Peltier cooling|
|Cooling temperature||Forced air (Ambient at +20 ℃): -10 ℃
Water (+20 ℃): -20 ℃
Water (+15 ℃): -30 ℃
|Dark current||0.06 electrons/pixel/s (-10 ℃) (typ.)
0.02 electrons/pixel/s (-20 ℃) (typ.)
0.006 electrons/pixel/s (-30 ℃) (typ.)
|Dynamic range||33 000:1 (typ.)*2|
|External trigger mode||Edge, Level, Synchronous readout and Start trigger|
|External trigger signal routing||SMA connector or CameraLink I/F|
|Trigger delay function||0 μs to 10 s in 10 μs steps|
|Trigger output||3 programmable timing outputs
Global exposure timing and Trigger ready output
|External signal output routing||SMA connector|
|Interface||CameraLink full configuration Deca mode / USB 3.0|
|Software interface||PC-based acquisition package included
DCAM-SDK, commercially available software
|A/D converter||16 bit output*3|
|Power supply||AC100 V to AC240 V, 50 Hz/60 Hz|
|Power consumption||Approx. 70 VA|
*2 Full well capacity / Readout noise in slow scan
*3 The true 16 bit image data is achieved through seamless merging of the output from two 11 bit A/D converters.
*Readout speed at center position (frames/s, typ.)
|Camera Link||USB 3.0|
|2048 / 1536 / 1024 / 512||2048 / 1536 / 1024||512|
|8||25 655||7894||25 655||25 655|
|Standard scan (at 100 frames/s)||10 ms|
|Slow scan (at 30 frames/s)||33 ms|
Lightsheet Readout Mode™ (Camera Link only)
|Readout format||Seamless readout|
|Readout direction||Top to bottom / Bottom to top|
|Readout time||20 ms to 204.8 s (at full area readout)|
|Scan mode||Full area, Sub-array|
Instruction manual [2 MB/PDF］
ORCA-Flash4.0 White paper [1.4MB/PDF]
Digital Cameras for Scientific Imaging [0.5MB/PDF]