Photometrics QuantEM:512SC Frequently Asked Questions

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1 Photometrics QuantEM:512SC Frequently Asked Questions What s new about the QuantEM camera? The new QuantEM:512SC camera from Photometrics offers: Linearized and quantitative EM gain slider provides a much more intuitive, easily quantifiable EMCCD camera. Self-calibrating EM gain feature ensures that the camera provides the EM gain requested. Due to the impactionization process, the EM gain can decline with use of the device. This selfcalibration feature allows the camera to remain quantitative over time. PAR (Photometrics Active Regulation) feedback system ensures that the image signal remains tightly controlled and measurement errors are minimized when using EM mode. This feedback system enhances the quantitative nature of the camera. patent pending ACE (Advanced Clocking Enhancement) technology increases Photometrics ability to control the voltage-clock timing resolution by 12x, effectively enabling the camera to achieve the lowest possible noise floor. This technology results in vastly improved quantitative ability and image quality. patent pending Stabilized background (bias stability) enabled by intelligent FPGA design allows a constant background with no drift during streaming, time lapse, etc. This critical advance permits far more quantitative data to be obtained for ratiometric and time-course experiments (e.g., FRET, Fura). 16-bit A/D conversion on the EM port and extremely low noise provide a high signal-to-noise ratio, which is especially important for low-light fluorescence imaging. With high QE and very low read noise, the new QuantEM:512SC camera is perfect for demanding microscopy applications. Turbo 1394 is Photometrics unique implementation of the IEEE 1394a (FireWire) interface. The new QuantEM:512SC provides the fastest parameter-switching image acquisition available on the market. This capability is vital for applications that require two or more exposures with different imaging parameters (e.g., high-speed ratio imaging applications).

2 What does the SC stand for in QuantEM:512SC? S stands for stabilized. This refers to bias and gain stability. C stands for cooled. This refers to the -30 C cooling. What does the stability of the QuantEM:512SC camera mean for my research? You can acquire repeatable data sets with this EMCCD camera. The stable bias (offset) provides a constant background for your imaging. This means that areas with no light on the sensor will remain at the same mean intensity during time lapse, data streams, etc. The stable gain means that the EM register amplification of the signal remains stable each time you use the detector (i.e., image to image, day to day, month to month, year to year). Why should I buy a QuantEM:512SC camera? The QuantEM:512SC camera has been designed specifically for low-light quantitative fluorescence microscopy applications in the life sciences. If you are doing live- or fixed-cell fluorescence imaging with extremely low light levels and short exposure times (<1 sec), then this EMCCD camera is the perfect choice for your work. The QuantEM:512SC camera is particularly well suited for the following applications: Widefield confocal microscopy Ratiometric imaging (e.g., Fura) Ratiometric FRET imaging of live cells FRAP imaging 4D imaging

3 How does the QuantEM:512SC camera s Turbo 1394 interface help me? This unique implementation of the IEEE 1394a interface provides two major benefits: Very high sustained frame rate while switching parameters. Normally when a parameter (e.g., exposure time or gain) is changed, it is necessary to reload a script to the camera. By minimizing the amount of new information sent to the camera, the frame rate stays higher. Very robust data transfer. Turbo 1394 transfers all frame data for each and every frame. It does not drop frames and does not give black frames. Every frame is a good frame, so you always get all the data you requested. What is the bit depth of the QuantEM:512SC camera? The new QuantEM:512SC is a 16-bit camera, thus it delivers much better fidelity than a 14-bit camera. The QuantEM:512SC camera s ability to measure smaller signals with more precision provides both excellent image quality and highly quantitative data. Can I use the QuantEM:512SC camera to trigger another piece of hardware? Yes, you can. The QuantEM:512SC camera affords extensive TTL-level functionality. This feature allows you to set up a wide range of simple and more complex hardware-triggered experiments. Do I have to use the QuantEM:512SC camera s external power supply? Yes, you do. The camera draws all its electricity from the external power supply. The camera s power supply automatically senses the incoming AC voltage. A common IEC 320 power cord receptacle is included for worldwide compatibility. Please note that the camera does not draw power from the computer.

4 Does the order in which I connect the power supply and data cable matter? No, but all cables must be connected before applying power to the camera. Do not disconnect the power supply while the camera is in use (e.g., being used for imaging, transferring data, and so forth). If the power supply is disconnected while the camera is in use, the camera power will be interrupted. It will then be necessary to disconnect (unplug) and reconnect (re-plug) the camera to restore functionality. What is the longest cable I can use with the QuantEM:512SC camera? The upper limit is 4.5 meters (about feet). A 3.0-meter cable (9.84 feet) is supplied with the QuantEM:512SC. The camera should be compatible with IEEE 1394a repeaters, but Photometrics does not guarantee camera performance with repeaters. Can I change the temperature of the QuantEM:512SC camera? Yes, you can. The temperature is set to -30 C, but can be changed by the user. The QuantEM:512SC camera will attempt to operate at the desired temperature, but may or may not achieve it depending on the temperature requested. My experiment is extremely sensitive to vibration. What can I do? You can use a remote fan to cool the camera s EMCCD. Contact your dealer for details. I received an IEEE 1394a board with my QuantEM:512SC camera. Do I have to use it? Photometrics recommends that you use the supplied board to ensure proper functionality. Built-in IEEE 1394a ports or other expansion boards may work, but are not guaranteed to do so. Which operating systems are supported by the QuantEM:512SC camera? The QuantEM:512SC camera is compatible with Windows XP (Service Pack 2 or later), Windows 2000 (Service Pack 4 or later), and Mac OS X (Power PC, version 10.3 or later).

5 Which imaging software can I use with the QuantEM:512SC camera? The QuantEM:512SC camera is compatible with many third-party imaging applications. Ask your dealer to supply a PVCAM -compatible imaging application to ensure support. The camera also comes with Saguaro software for Windows, a program that allows full camera functionality as well as image capture and analysis. What is the QuantEM:512SC camera warranty? One (1) year on parts, labor, and manufacturing defects What comes with the QuantEM:512SC camera? Camera Test report Quick Installation guide Data cable (Turbo 1394: 6-pin to 6-pin connectors, 3.0-meter length) Brick-style power supply with power cord Interface board (Turbo 1394: IEEE 1394a board with 6-pin to 6-pin connectors) C-mount, fixed-aperture lens Lens cap Registration card Software CD-ROM, including: o RS Image software for Mac and Windows o Saguaro image capture and analysis software for Windows o Small software module for EM gain calibration o PVCAM drivers o Printable product manuals (.pdf files)