vitrobot
Differences
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| Both sides previous revisionPrevious revisionNext revision | Previous revision | ||
| vitrobot [2023/01/13 14:32] – crucifix | vitrobot [2023/11/01 20:18] (current) – external edit 127.0.0.1 | ||
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| * fill a dry 5 L tank with liquid nitrogen | * fill a dry 5 L tank with liquid nitrogen | ||
| * label grid boxes with the name of the experiment | * label grid boxes with the name of the experiment | ||
| - | * check several things: | + | * check plasma cleaner is reserved |
| - | | + | * Assemble the necessary tools: |
| * EM_grids grids are available and present in sufficient number | * EM_grids grids are available and present in sufficient number | ||
| - | * all part of the coolant container are dry | + | * all part of the coolant container |
| - | * both vitrobot tweezers, big tweezer and blot paper clipping | + | * both vitrobot tweezers, big tweezer and blot paper clipping |
| ==== Starting up the vitrobot ==== | ==== Starting up the vitrobot ==== | ||
| Line 56: | Line 56: | ||
| * 1. 'blot total' needs to be ' | * 1. 'blot total' needs to be ' | ||
| * 2. check ' | * 2. check ' | ||
| - | * 3. set your desired blot time, force blot and waiting time (o for unsupported condition ) | + | * 3. Optional : set your desired blot time, force blot and waiting time (o for unsupported condition ) |
| + | * all these setting are checked at the begining of the freezing cycle | ||
| | | ||
| ==== Set humidity and temperature parameters in climate chamber ==== | ==== Set humidity and temperature parameters in climate chamber ==== | ||
| Line 123: | Line 124: | ||
| * the metallic drawer is **ALWAYS stored in the chamber** | * the metallic drawer is **ALWAYS stored in the chamber** | ||
| * 5. close the door to prevent dust contamination inside the chamber | * 5. close the door to prevent dust contamination inside the chamber | ||
| - | * 6. ++ place EM_grids inside the metallic drawer |< | + | * 6. place EM_grids inside the metallic drawer |
| + | * ++ EM_grids with support film facing up|< | ||
| * remove the cover of the metallic drawer by sliding | * remove the cover of the metallic drawer by sliding | ||
| * place EM_grids inside | * place EM_grids inside | ||
| Line 149: | Line 151: | ||
| * 11. if not yet done on vitrobot, enable the humidity switchbox to start | * 11. if not yet done on vitrobot, enable the humidity switchbox to start | ||
| - | ==== Assemble different part of the coolant container | + | ==== Assemble different part of the coolant container |
| {{ : | {{ : | ||
| Line 244: | Line 246: | ||
| - | + | ======Everything | |
| - | + | ||
| - | + | ||
| - | ==== Start freezing at the vitrobot | + | |
| - | + | ||
| - | + | ||
| - | * 1. check the blotting condition | + | |
| - | * 1. click on "place new grid" to bring in loading position the plunge rod | + | |
| - | * 2. Attach Vitrobot tweezers with a Glow discharged grid onto the plunge rod | + | |
| - | * 3. press to bring up the tweezers into the blotting chamber | + | |
| - | * 4. place the container onto the ethane lift | + | |
| - | * 5. press to lift it | + | |
| - | * 6. click on " | + | |
| - | * 7. apply sample to grid | + | |
| - | * 8. click on " | + | |
| - | * 9. carrefully detach tweezers from the plunge rod | + | |
| - | * 10. By maintaining the grid inside the ethan cup, place the coolant container on the table | + | |
| - | * 11. transfer grid to the grid position | + | |
| - | * 12. Dry tweezers then repeat | + | |
| - | + | ||
| - | * Now vitrobot | + | |
| - | + | ||
| - | ====== | + | |
| - | + | ||
| - | At this point already is ready for freezing: | + | |
| climate chamber is equilibrated in temperature and humidity. | climate chamber is equilibrated in temperature and humidity. | ||
| - | Filter paper are mounted | + | Filter paper are mounted |
| ethane is liquefied | ethane is liquefied | ||
| - | Grids are loaded and ready to be plasma cleaned | + | Grids are freshly |
| - | | + | ==== Start freezing at the vitrobot ==== |
| - | ==== Preparation of the grid ==== | ||
| + | * 1. check that the correct freezing parameters are entered | ||
| + | * 2. click on "place new grid" to bring in loading position the plunge rod | ||
| + | * 3. Lock a Glow discharged grid on tweezers | ||
| + | * Grasp as little of the grid as possible with the tweezers (but enough that it will not fall out during freezing) | ||
| + | * ++ Make sure that the black clamping ring is fixed not more down to the first groves. |< | ||
| + | {{ : | ||
| + | </ | ||
| + | * 4. then fix the tweezers onto the plunge rod | ||
| + | * with the support film support on the right for right-handers and on the left for left-handers | ||
| + | * 5. press on " | ||
| + | * 6. place the container onto the ethane lift | ||
| + | * Always check that ethane is at right temperature | ||
| + | * 7. press on to lift it | ||
| + | * 8. click on " | ||
| + | * 9. apply sample to grid | ||
| + | * 10. click on " | ||
| + | * 11. carrefully detach tweezers from the plunge rod | ||
| + | * 12. By maintaining the grid inside the ethan cup, place the coolant container on the table | ||
| + | * 13. transfer grid to the grid position | ||
| + | * 14. Dry tweezers | ||
| + | * 15. then repeat sequence 1-14 for all glow discharged grids | ||
| - | ++3. Bring the vitrobot to hermetically close the chamber|< | ||
| - | * do the following dry Run: | ||
| - | * A glow discharge grids is attached to the twezzers | ||
| - | |||
| - | ==== Start freezing ==== | ||
| - | |||
| - | | ||
| - | * ++ Do it when you are almost ready to freeze to keep the soaking time as reproducible as possible|< | ||
| - | Decrire | ||
| - | </ | ||
| - | |||
| - | ==== Mounting the filter papers, IMPERATIVELY wear gloves ==== | ||
| - | |||
| - | * ++ Do it when you are almost ready to freeze to keep the soaking time as reproducible as possible|< | ||
| - | Decrire | ||
| - | </ | ||
vitrobot.1673620331.txt.gz · Last modified: (external edit)