C11440-22CUORCA-Flash4.0 V2 Digital CMOS camera
It's here! The new QE-enhanced ORCA-Flash4.0 V2 with 82 % peak QE.Scientific breakthroughs rarely come from giant steps. Rather, it’s a continuous progression of small steps and astute application of those differences that enables advances. The ORCA-Flash4.0 V2 is the perfect examples of this process. From its introduction the ORCA-Flash4.0 has challenged the status quo of imaging and undergone a series of useful enhancements. The most recent is perhaps the most exciting; a notable increase in the ability to detect photons. This enhanced QE means that you have a possibility of detecting the faintest of signals. Or, for moderately dim samples, that the ideal image is achievable with shorter exposure times, perhaps saving your cells from phototoxicity or bleaching. With the ORCA-Flash4.0 V2 already delivering wide field of view, large dynamic range, and fast frame rates, this QE enhancement only makes it more versatile and powerful.
If you have not yet experienced the ORCA-Flash4.0 V2 sCMOS, now is the time.
What breakthrough will you make with your extra photons?
Enhanced Quantum Efficiency
The QE is the wavelength dependent probability that a photon is converted to a photoelectron.
The enhanced QE of the ORCA-Flash4.0 V2 is 82 % peak QE.
Two Scan Speeds
While the read noise at standard scan is only 1.6 electrons rms (1.0 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 1.4 electrons rms (0.8 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!
Does a little bit more Quantum Efficiency (QE) make a difference?
An easy way to consider this question is to compare two cameras with identical specs except QE. Both cameras have very low read noise of 1.4 e- rms. But Camera A has 82 % QE at 560 nm and Camera B has 72 %. Does this 10% matter? At low light, this higher QE reduces the impact of the read noise. It sounds counterintuitive, but here’s the logic.
Generally, read noise for cameras is specified in electrons, which is wavelength independent. But light is the thing we are measuring and we believe it should be reported in photons. It is the relationship of the camera noise to the QE at the wavelength of interest that holds the key to understanding why the effective read noise is different in the two cases described. The math is simple. For Camera A, this would translate into 1.4 e-/.82 or 1.7 photons and for Camera B its 1.9 photons. That’s actually a 14% higher read noise in Camera B and this difference can be relevant for very low light imaging. The reality is that many biological samples enough are bright enough for this read noise difference to be inconsequential. But at these low-to-mid intensities, where the answers to many complex biological questions exist, the higher QE contributes to better overall signal to noise and provides the practical capability to increase frame rates, reduce illumination intensity or shorten exposures without sacrificing SNR.
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 1.0 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 1.6 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.
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: 82 % peak QE
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 : 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. 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 1.6 electrons rms (1.0 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.
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, 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.
PC Recommendations for ORCA-Flash4.0 [747 KB/PDF］
|Quantum efficency||82 % peak QE|
|Imaging device||Scientific CMOS Sensor|
|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.):1.6 electrons rms (1.0 electrons median)
Slow scan (at 30 frames/s, typ.): 1.4 electrons rms (0.8 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||37 000:1 (typ.)*2|
|External trigger mode||Edge, Level, Synchronous readout, Start trigger, Global reset edge and Global reset level|
|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||Camera Link*3 / USB 3.0|
|Software interface||PC-based acquisition package included
DCAM-SDK, commercially available software
|A/D converter||16 bit output*4|
|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 Proprietary mode equivalent of Camera Link 80-bit configuration
*4 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||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]