Safety considerations

The Setup operates with high currents and voltages and is therefore dangerous for the operator and other instruments. Dangers include but are not limited to:

• Electrical discharged
• Heat / Cold burns from the Peltiers
• Water damage in case of leakage

Whenever anything on the setup is changed, make sure to check the proper operation of the setup.

Preparation

Before any usage of the setup it should be checked that it is working as inteded (off the microscope).

Check before every usage:

• Wire connections
• Thermistor placement
• Tightness of all tube conenctions and Peltier - Backplate connection

Do never power the Arduino through the USB connection when the power switch is turned ON, as this can damage the Arduino and driver boards.

Motor calibration

Since the motor can be moved when the setup is powered off it can happen that the motorposition on the Arduino is not accurate. If the motor drives to the end of the travel, it is possible that the motor damages itself. Therefore it is important to check the motor calibration every time the motor is used. To do this carefully drive the motor to the zero position and check, wether it is the real zero position as indicated by the alignement on the motor and the backplate. If this is not the case, move the motor to the real middle position and re-calibrate the zero position by pressing Setting/Calibrate. The motor can for this also be moved by disabling Settings/motor hold and turning the barrel on the opposite end of the motor.

Condensation prevention

Humidity can be a large problem for the experiment, either because it makes imaging of the sample impossible or it can lead to frost on the sample movement frame and prevent sample translation. Therefore it is important to check and close all places where humidity could enter the setup. It is good practice to put a glass slide with dessiccator beads on it onto the copper block during the experiment. Furthermore any condensation should be removed before the Setup is closed. After the experiment it should be studied where frost / condensation appeared in the setup and the enclosure should be optimised accordingly for the next experiment

Standard operation protocol (SOP)

1. Remove neighboring objectives from Objetive turret; add Obejctive range extender (2x 15mm)
2. Test wet-power-shutoff Sensors by touching that they turn the power off (will beep when succesfull)
3. Mount spil-protection with Water sensors
4. Add 3D printed frame with plastic against humidity
5. Connect all cables of Gelatiera 3 to power, check all wires and tubes for damage
6. Run at a comparable temperature Profile to the target profile for at least 10 minutes off the microscope. Check for water leakage and cooling performance.
7. Choose and mount correct translational frame for your sample. The tighter the better
8. Heat stage head up to Room temperature or 35 °C to remove any condensation that appeared during the test run. Wait 10 minutes
9. Cool to room temperature, mount sample and if necessary Silica beads (trap humidity) next to sample on copper blocks and next to objective
10. Mount Gelatiera hat (top humidity protection)
11. Mount Stage head on carefully on microscope. Do not push on the piezo stage with this weight. Make sure the stage head is fixed and does not wobble in the frame
12. Apply the intended temperature profile. Slow cooling rates can help against condensation
13. Move the motor to the middle position (either digitaly or by turning Settings/Motor hold off and turning the barrel on the motor by hand) and reset the 0 position of the motor.

After the experiment

1. Remove stage head from microscope
2. Remove Gelatiera hat, observe where ice/condensation formed to improve performance for the next experiment
3. Remove sample
4. Dry stage head by heating to 35 °C
5. Disconnect power and store all cables/parts at their designated place

Troubleshooting

Sensors not working

• Tap all cable connection points to find loose contacts.
• put 10 or 30 kOhm resistor into various places where the sensor is connected (Control box, signal cable, stage head) to localized the broken connection.