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The NeuroCCD series of cameras is ideal for low and medium light level measurements. They are used to study a wide range of dye stained preparations: from filled single cells through preparations stained by transported dye to stained tissue slices.
The NeuroCCD-SM256 camera is ideal for Calcium, pH and intrinsic imaging measurements with very high spatial resolution.

256 x 256 pixels
Full-frame rate: 100 fps
3x3-binning rate: 270 fps
Read Noise
  100 Hz:
62 Hz:
38 Hz:
28 e-
15 e-
11 e-

Very large well size : 750,000 e- at full frame 1,000,000 e- at 2X2 binning
Frame shift time : 322 microsecs
Uses the outstanding Marconi CCD67 chip
Digitization: 14 bits standard, 16 bits optional (slower frame rate)
Back-Illuminated for high quantum efficiency (80% at 400-600nm, 75% at 650nm)
Cooled for low dark noise
No fan; avoids vibrations
Simultaneous optical and 8 channels of electrical recording
Optical coupler to reduce objective image to the size of the CCD chip
Comprehensive, user-friendly software designed for neurobiologists, physiologists and biophysicists
Computer control of shutter and stimulator.


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Optimal Use of the CCD Camera
The S/N ratio at moderate and high light levels is the square root of the light intensity. To increase the S/N one should pump in (and collect) as much light as possible at the lowest gain (1X). Only at this gain (1X) one uses the full well-size of the CCD and this is the preferred mode of measurement for most bath-stained tissue, such as brain slices and cardiac preparations. The saturation at higher gains (>=3X) is probably amplifier saturation. If saturation occurs at 1X, try to use a higher frame rate (100Hz, unbinned). This will allow emptying the CCD wells faster, thus avoid saturation w/o losing light. One can temporally bin the data later to increase the S/N ratio. If saturation still occurs when using the highest frame rate, one will have to reduce the illumination. Higher gains should be used at dim light levels to better utilize the 14-bit digitization.

Computer
We strongly recommend reserving the computer which comes with NeuroCCD-SM256 for this function only. The reasons are two-fold. First, data acquisition places large demands on the computer's resources (22 Mbytes per second data transfer at 2.0 kHz frame rate). Second, it is very difficult for RedShirtImaging to troubleshoot any problems that might arise when attempts are made to add additional hardware and software. Two examples follow:

First, we installed an Imagenation PX-500-10 frame grabber in the same computer with NeuroCCD-SMQ. This caused NeuroCCD-SMQ to miss frames at 1.0 kHz. We presume that this is a solvable problem (but not with our level of expertise at PC administration).

Second, a user tried to install a SCSI card. This generated a conflict with the a-to-d interrupt request and the SCSI installation did not work. Then uninstalling the SCSI left the a-to-d card non-functional.



Shutter
A shutter which can be driven by a 5-volt pulse should be available; shutter opens at 5 volts, closes at 0 volts. In addition to limiting the amount of photo-toxicity, the use of a shutter is essential for some frequently used features of the software. We use shutter drivers and shutters from Vincent Associates. Choose a shutter opening that is larger than the diameter of the incident light beam. We mount the shutter independently of the microscope so that its vibration does not affect the recording.

IDL licensing
The NeuroPlex software is written on an IDL platform. A full IDL license is supplied with our cameras allowing the end user to modify or add functionality. We encourage users who are interested in modifying the software to contact us first.

IGOR Pro/Visual Basic drivers
The IGOR/Visual Basic drivers for the NeuroCCD are recommended for labs that already use IGOR or Visual Basic for imaging and thereby have experience using these languages for driving a camera and have routines for display and analysis of the optical signals.

For labs that do not have this experience and the display-analysis routines, we expect that it will take a number of months to learn how to drive the camera and to develop the initial routines. For these labs we recommend purchase of the full NeuroCCD system together with an IGOR/Visual Basic license (an additional $600). In this way the camera can be used for experiments immediately while the IGOR/Visual Basic programs are developed. The NeuroCCD software can also be helpful by providing a comparison for debugging the IGOR/Visual Basic software.

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