3. BNC male to terminal block adapter (e.g., Delock No. 65323,
Tragant, Berlin, Germany) (see Note 15).
4. BNC “T” splitter, three-way female adapter.
2.5 Tools
1. Handheld blow-dryer.
2. Bulb air blower (e.g., Rocket-air AA 1900, Giotto’s Industrial,
Taipei, Taiwan).
3. Two pairs of fine-pointed forceps, angled (e.g., Fisherbrand
No. 08-875, Thermo Fisher Scientific, Waltham,
Massachusetts, USA).
4. Adjustable micropipette, 1–10 μL, with disposable tip.
5. Vacuum tweezers (e.g., PEN-VAC Pro Series, Virtual Industries, Colorado Springs, Colorado, USA) (see Note 16).
6. Stopwatch.
7. Flashlight.
2.6 Materials
and Supplies
1. Syringe, 5 mL, Luer-lock.
2. Blunt-tipped dispensing needle, 16 G, Luer-lock.
3. Silicone grease (e.g., High Vacuum Grease, Dow Corning,
Midland, MI, USA).
4. Isopropyl alcohol.
5. Glass coverslips, circular, 16-mm diameter (e.g., Part
No. W16G, Linkam Scientific Instruments) (see Note 17).
6. Low-lint delicate task wipers (e.g., Kimtech Science Kimwipes
No. 34155, Kimberly-Clark, Roswell, Georgia, USA).
7. Liquid nitrogen.
8. Cell suspension, approximately 10
6 cells/mL (see Note 18).
3 Methods
3.1 Trigger
Interface Cable
To obtain accurate kinetic data, it is necessary to synchronize as
closely as possible the independent timing signals (i.e., the clocks)
of the cryomicroscope system controller and the high-speed imaging system, which requires a channel of communication between
the two systems. For versions of the Linkam Scientific cryomicroscopy systems available since 2016 (which use the LINK software),
it is straightforward to establish communication between the cryomicroscope control unit (model T95 or T96) and a third-party
high-speed video camera, via the controller’s input port for TTL
signals. To synchronize the camera clock with the cryomicroscope
system clock, a trigger interface cable connecting the camera and
the cryomicroscope system controller can be constructed as shown
in Fig. 1a (see Note 19).
High-Speed Video Cryomicroscopy
225
Tragant, Berlin, Germany) (see Note 15).
4. BNC “T” splitter, three-way female adapter.
2.5 Tools
1. Handheld blow-dryer.
2. Bulb air blower (e.g., Rocket-air AA 1900, Giotto’s Industrial,
Taipei, Taiwan).
3. Two pairs of fine-pointed forceps, angled (e.g., Fisherbrand
No. 08-875, Thermo Fisher Scientific, Waltham,
Massachusetts, USA).
4. Adjustable micropipette, 1–10 μL, with disposable tip.
5. Vacuum tweezers (e.g., PEN-VAC Pro Series, Virtual Industries, Colorado Springs, Colorado, USA) (see Note 16).
6. Stopwatch.
7. Flashlight.
2.6 Materials
and Supplies
1. Syringe, 5 mL, Luer-lock.
2. Blunt-tipped dispensing needle, 16 G, Luer-lock.
3. Silicone grease (e.g., High Vacuum Grease, Dow Corning,
Midland, MI, USA).
4. Isopropyl alcohol.
5. Glass coverslips, circular, 16-mm diameter (e.g., Part
No. W16G, Linkam Scientific Instruments) (see Note 17).
6. Low-lint delicate task wipers (e.g., Kimtech Science Kimwipes
No. 34155, Kimberly-Clark, Roswell, Georgia, USA).
7. Liquid nitrogen.
8. Cell suspension, approximately 10
6 cells/mL (see Note 18).
3 Methods
3.1 Trigger
Interface Cable
To obtain accurate kinetic data, it is necessary to synchronize as
closely as possible the independent timing signals (i.e., the clocks)
of the cryomicroscope system controller and the high-speed imaging system, which requires a channel of communication between
the two systems. For versions of the Linkam Scientific cryomicroscopy systems available since 2016 (which use the LINK software),
it is straightforward to establish communication between the cryomicroscope control unit (model T95 or T96) and a third-party
high-speed video camera, via the controller’s input port for TTL
signals. To synchronize the camera clock with the cryomicroscope
system clock, a trigger interface cable connecting the camera and
the cryomicroscope system controller can be constructed as shown
in Fig. 1a (see Note 19).
High-Speed Video Cryomicroscopy
225