Leica EM TP User notes Leica EM TP (Tissue Processor) General operation 1. Switch on at the back Sealing rings are on the lid, once they are worn out, turn them over. Need to have a supply of consumables. Bright red lids for capping fixatives etc. 2. Fix in the hood as per usual. 3. Set up the solutions in the EMTP whilst fixation is being undertaken. Don’t user over 15 ml, especially for resins. You may need to overfill to compensate for ethanol evaporation. 4. 24 places for the samples. (1) is the position for the samples to be placed in initially. The inner numbers are for the light microscopy baskets, which we do not have the set-up for. 5. Tighten the holder with all the samples in the EM TP. The knob should screw on 3-4 times, if it tightens with only ½ a turn, the holder is not level and will cause problems during the run. 6. The stand is where you put the baskets in order. Deeper holes are for the 3 division baskets. 7 baskets, up to 56 samples. Make a note of your samples either on the specialised sheets or use small pieces of paper in each basket. Numbers are from 1-7. Have the final holder filled with buffer. 7. When samples are sorted out, pick each basket up from 1-7 using the metal rod. Either put the clip on top before the baskets are picked up or after. Wind the rod onto the base quickly and place in the buffer. 8. Clip the rod into its EM TP position, close the lid. 9. Select program and press start. If you want it to finish in the morning, change the finish time to when you want to finish it. 10. At the end of the program, unhook the rod and samples from the holder, bring the vials down and pull out the final vial and take to the fume cupboard for embedding. Bioimaging @Rothamsted Research Leica EM TP User notes Programming 1. Buttons: Program - Input Key – Locks the keyboard, hold down for 3-4 seconds. Lower right hand buttons - for manual use (circle to move the vials around, up arrow to raise the vials up, down arrow to bring the vials down, agitation for agitating, temperature for changing temperature). Cooling by peltier cooling, heating by element. 2. To program: 3. Press program. 01 – Title – e.g., LRW – LM, hold key down to get to correct number or letter. Across arrow for next space. EM always chosen for our system with 24 vials Vial 1 – Buffer (003) Agitation – 03 (standard) Vial 2 – Buffer (003) Agitation – 03 (standard) etc. Vial 3 – Distilled water etc. Last resin set to 6 hours. Press program again to finish and program again to save. **Battery backup in case of power failure. It will turn off any cooling or heating and finish the run. If it happens just at the end of the sample run, it might restart the run. Always put last resin for 6 hours. Can input programs through a laptop. Faster and quicker. Up to 99 programs can be put in. Bioimaging @Rothamsted Research