diff --git a/prep/vitrobot.rst b/prep/vitrobot.rst index ed9cbb0..299b4db 100644 --- a/prep/vitrobot.rst +++ b/prep/vitrobot.rst @@ -68,13 +68,17 @@ Material Preparation * Prepare Falcon tubes, grid boxes and screwdrivers or rods * **Prepare the coolant container**: - * Insert brass coloured inner cup, grid box holder, the spindle/spider and anti-contamination ring + * Insert brass coloured inner cup, grid box holder, the spindle/spider and anti-contamination ring (figure 1) * Fill up the inner (brass) and outer chamber (styrofoam) with LN2 and let it settle under a glass cupola (to prevent contaminations from humidity in the air) * Wait until LN2 evaporates from the brass coloured cup (should take around 5 minutes), top up the outer chamber if necessary + | + | + .. image:: /images/Coolant_chamber.png + | * **Condense ethane:** * **WEAR GLASSES AND GLOVES, ALWAYS WORK IN THE FUME HOOD**, make sure that all LN2 have evaporated from the ethane cup - * Open the valve 1, and valve 3 on the on the ethane bottle (figure 1) + * Open the valve 1, and valve 3 on the on the ethane bottle (figure 2) * **Carefully** open valve 2 of the ethane bottle, aim for your skin on the wrist above you gloved hand. When you can feel a gentle stream of gas on your skin, the inital stream of gas i sufficient (on the manometer the pressure should be around 1). **Better too slow than too fast!** * Close valve 2. Put the pipette tip of the out-flow at the bottom of the ethane cup, **quickly** turn back the flow of ethane by turning valve 2 back to the position of the gentle stream. * Start filling the ethane cup with ethane. After 10-30 seconds the sound of the ethane should start to zizzle (as a result of liquid ethane forming) @@ -87,9 +91,10 @@ Material Preparation * Top up the LN2 as required * Place your marked grid box(es) onto the grid box holder (TEST them!) * Transfer the coolant chamber and LN2 thermos flask next to the Vitrobot - + | + | .. image:: /images/Valves_ethane.png - + | * **(Condense ethane with a NanoSoft ethane-condenser)**: * **WEAR GLASSES AND GLOVES, ALWAYS WORK IN THE FUME HOOD**, make sure that all LN2 have evaporated from the ethane cup * Follow the instructions on the following video: https://youtu.be/J_M-IUlPuNM