system must incorporate a circular buffer architecture with at
least 1 GB of memory (preferably 10 GB or more) and the
ability to configure the timing of the trigger event that controls
termination of image acquisition; the camera must accept an
external trigger signal for this purpose. In addition, the frame
rate, image resolution, and exposure time should be useradjustable. The camera’s light sensitivity and signal-to-noise
ratio should be sufficient to yield publication-quality micrographs at exposure times (i.e., shutter speeds) of 100 μs; this
must be confirmed by imaging tests using the actual cryomicroscope setup on which experiments will be performed,
including a representative cell sample.
3. High-speed imaging systems typically include a dedicated software package for configuration and control of the camera.
Older Vision Research Phantom camera models, such as the
v7.1 model used here, require a pre-2010 implementation of
the Vision Research Phantom Camera Control application,
which is referred to as the “legacy” version of the software. In
contrast, more modern Phantom camera models use a post2010 version of the Vision Research Phantom Camera Control
program, which is generally referred to as the “PCC” software.
In this chapter, distinct instructions are provided for both
versions of the software.
4. The capture bundle cable is a breakout cable that provides
access to the camera’s hardware trigger input (as well as other
signal connectors), typically via a BNC connector. The required
capture cable hardware may be different for different camera
models. In the Vision Research Phantom v7.1 camera used
here, the capture cable joins a 19-pin Amphenol military standard circular connector to five BNC male connectors (which
correspond to the camera’s trigger, F-sync, pre-trigger, IRIGin, and analog video signals, respectively); this capture bundle
cable is supplied with the v7.1 camera system. In the most
recent high-speed imaging cameras from Vision Research, the
trigger signal connector is available directly on the back of the
camera chassis via a BNC female connector; for these cameras, a
standard BNC male to BNC male coaxial cable can be used in
place of the capture bundle cable.
5. The microscope used here is a Leica DM2500 M stand with a
5Â/0.12 NA, 14.0-mm working distance objective (Part
No. 11566077), a 20Â/0.40 NA, 6.9-mm working distance
objective (Part No. 11506242), as well as a universal condenser
(Part No. 11551062) with a 0.50-NA, 15-mm working distance top lens (Part No. 11501037); the microscope system
also includes a 100-W lamp housing (Part No. 11504080), a
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Jens O. M. Karlsson
least 1 GB of memory (preferably 10 GB or more) and the
ability to configure the timing of the trigger event that controls
termination of image acquisition; the camera must accept an
external trigger signal for this purpose. In addition, the frame
rate, image resolution, and exposure time should be useradjustable. The camera’s light sensitivity and signal-to-noise
ratio should be sufficient to yield publication-quality micrographs at exposure times (i.e., shutter speeds) of 100 μs; this
must be confirmed by imaging tests using the actual cryomicroscope setup on which experiments will be performed,
including a representative cell sample.
3. High-speed imaging systems typically include a dedicated software package for configuration and control of the camera.
Older Vision Research Phantom camera models, such as the
v7.1 model used here, require a pre-2010 implementation of
the Vision Research Phantom Camera Control application,
which is referred to as the “legacy” version of the software. In
contrast, more modern Phantom camera models use a post2010 version of the Vision Research Phantom Camera Control
program, which is generally referred to as the “PCC” software.
In this chapter, distinct instructions are provided for both
versions of the software.
4. The capture bundle cable is a breakout cable that provides
access to the camera’s hardware trigger input (as well as other
signal connectors), typically via a BNC connector. The required
capture cable hardware may be different for different camera
models. In the Vision Research Phantom v7.1 camera used
here, the capture cable joins a 19-pin Amphenol military standard circular connector to five BNC male connectors (which
correspond to the camera’s trigger, F-sync, pre-trigger, IRIGin, and analog video signals, respectively); this capture bundle
cable is supplied with the v7.1 camera system. In the most
recent high-speed imaging cameras from Vision Research, the
trigger signal connector is available directly on the back of the
camera chassis via a BNC female connector; for these cameras, a
standard BNC male to BNC male coaxial cable can be used in
place of the capture bundle cable.
5. The microscope used here is a Leica DM2500 M stand with a
5Â/0.12 NA, 14.0-mm working distance objective (Part
No. 11566077), a 20Â/0.40 NA, 6.9-mm working distance
objective (Part No. 11506242), as well as a universal condenser
(Part No. 11551062) with a 0.50-NA, 15-mm working distance top lens (Part No. 11501037); the microscope system
also includes a 100-W lamp housing (Part No. 11504080), a
244
Jens O. M. Karlsson