However, the high-speed video cryomicroscope design documented in this chapter is based on a pre-2016 Linkam Scientific
control system using the Linksys32 software, which is not capable
of communication via TTL signal ports on the system controller.
To circumvent this limitation, the solution described here takes
advantage of the availability of a dedicated signal channel used for
sampling pressure gauge measurements in system controllers that
are designed for vacuum-capable thermal stages produced by Linkam Scientific (see Notes 20 and 21). Thus, the trigger interface
cable required for this solution, shown in Fig. 1b, should be constructed as described below (see Note 22).
1. Connect the black (signal ground) wire from the 9-pin vacuum
gauge connector (pin 1) to the outer shield (ground, or negative terminal) of a BNC male connector, and connect the red
(signal) wire from pin 6 of the 9-pin connector to the central
tip (signal, or positive terminal) of the BNC connector (see
Note 23).
2. Using a three-way BNC female “T” adapter, create a trigger
interface cable by joining the BNC male connectors of the
vacuum signal cable (from step 1 above), the camera’s trigger
signal cable, as well as the trigger switch cable.
The three cables of the trigger interface cable assembly will
interconnect the system components as follows: one cable leads
to the connector that will be mated to the cryomicroscope
system controller unit (see step 4 in Subheading 3.2); another
cable leads to a connector that will be joined to the camera
system trigger input socket (see step 2 in Subheading 3.3); the
third cable is terminated by the pushbutton switch in the
manual trigger.
3.2 Instrumentation Setup
An overview of the complete high-speed video cryomicroscopy
system is shown in Fig. 2. This section provides instructions for
setting up the main components of the system, except for those
necessary to supply the liquid nitrogen coolant (setup of the cooling system is outlined in Subheading 3.3). The instrumentation
setup steps described in this section should typically be performed
only once; in contrast, the experimental setup protocol described in
Subheading 3.3 must be completed at the start of each day of
experimentation.
1. If using an external light source (see Note 24), remove the
transmitted-light lamp housing from the microscope stand,
attach the collimating adapter to the illuminator mount, and
then couple the light source output to the microscope collimating adapter using a liquid light guide (see Note 25).
2. If not already preinstalled, install the Linkam Scientific vacuum
controller expansion card into Slot 1 on the motherboard
High-Speed Video Cryomicroscopy
227
control system using the Linksys32 software, which is not capable
of communication via TTL signal ports on the system controller.
To circumvent this limitation, the solution described here takes
advantage of the availability of a dedicated signal channel used for
sampling pressure gauge measurements in system controllers that
are designed for vacuum-capable thermal stages produced by Linkam Scientific (see Notes 20 and 21). Thus, the trigger interface
cable required for this solution, shown in Fig. 1b, should be constructed as described below (see Note 22).
1. Connect the black (signal ground) wire from the 9-pin vacuum
gauge connector (pin 1) to the outer shield (ground, or negative terminal) of a BNC male connector, and connect the red
(signal) wire from pin 6 of the 9-pin connector to the central
tip (signal, or positive terminal) of the BNC connector (see
Note 23).
2. Using a three-way BNC female “T” adapter, create a trigger
interface cable by joining the BNC male connectors of the
vacuum signal cable (from step 1 above), the camera’s trigger
signal cable, as well as the trigger switch cable.
The three cables of the trigger interface cable assembly will
interconnect the system components as follows: one cable leads
to the connector that will be mated to the cryomicroscope
system controller unit (see step 4 in Subheading 3.2); another
cable leads to a connector that will be joined to the camera
system trigger input socket (see step 2 in Subheading 3.3); the
third cable is terminated by the pushbutton switch in the
manual trigger.
3.2 Instrumentation Setup
An overview of the complete high-speed video cryomicroscopy
system is shown in Fig. 2. This section provides instructions for
setting up the main components of the system, except for those
necessary to supply the liquid nitrogen coolant (setup of the cooling system is outlined in Subheading 3.3). The instrumentation
setup steps described in this section should typically be performed
only once; in contrast, the experimental setup protocol described in
Subheading 3.3 must be completed at the start of each day of
experimentation.
1. If using an external light source (see Note 24), remove the
transmitted-light lamp housing from the microscope stand,
attach the collimating adapter to the illuminator mount, and
then couple the light source output to the microscope collimating adapter using a liquid light guide (see Note 25).
2. If not already preinstalled, install the Linkam Scientific vacuum
controller expansion card into Slot 1 on the motherboard
High-Speed Video Cryomicroscopy
227