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PERKIN ELMER ROBOTIC PIEZORRAY SYSTEM
General set-up for use.
Overview - The deposition of small volumes of liquid are subject to rapid evaporation, well in advance of conclusion of the array printing. Evaporation can be controlled by marinating the system in a state of balance where evaporation and condensation do not occur. This can be achieved by rigorous control of chamber humidity and plate temperature.
Humidity is controlled by via the black unit inside the enclosure on the far right. Humidity is a result of production of water vapour by this unit. To produce vapour it must be filled (to the port) with distilled water. Chamber humidity and air temperature are assessed by the Thermohygrometer.
Plate temperature is controlled by adjusting the chiller temperature, located outside the enclosure and to the right of the arrayer. This is a closed system and does not require water. Plate temperature is assessed by the Thermocouple.
Local weather and seasonal changes affect relative humidity in the chamber. Optimum spotting occurs when chamber humidity is around 40% relative humidity and the dew point is around 13C. Large differences in humidity between the chamber and lab make maintenance of this target difficult.
Daily Start-up List for Piezorray
1) Press stop button to end standby mode
2) Run Prime Utility. Perform steps 3,4,5 while priming
3) Check level in System Liquid bottle. (> ¼ bottle)
4) Check helium sparging for proper operation (tank has gas, bubbles are flowing, bubbles are away from lines) Flow should read 30-50, tank 100 on first dial 9-10 on second.
5) Turn on Chiller and allow time for temperature stabilization
6) Prepare Plate for deep cleaning (PBS + 2% Tween 20 in 96 well plate using wells A1, B1, C1, D1)
7) Perform deep cleaning utility (perform steps 8,9) Once deep cleaning command reaches the 96 well plate perform steps 10,11,12,13)
8) Clean and fill Buffer Trough (rinse in dH20) Fill with 14 ml 1X sterile PBS
9) Clean and fill Cleaning Trough (rinse in dH20 from tap) Fill with 14 ml 1X sterile PBS + 1% Tween.
10) Fill Wash reservoir with dH20 from tap
11) Empty Waste reservoir (sink)
12) Fill enclosure humidification reservoir with dH20. Turn on enclosure pump.
13) Prepare source plate for printing. (Centrifuge source plate for 5 minutes @ 1100g to ensure that all air bubbles have been displaced.
14) Perform prime utility
15) Perform Tip Tuning (tune the tips to your samples for optimal performance. Use the buffer only it is an adequate representation of the protein solution)
Daily Start-up List for Piezorray
1) Press stop button to end standby mode
2) Run Prime Utility. Perform steps 3,4,5 while priming
3) Check level in System Liquid bottle. (> ¼ bottle)
4) Check helium sparging for proper operation (tank has gas, bubbles are flowing, bubbles are away from lines) Flow should read 30-50, tank 100 on first dial 9-10 on second.
5) Turn on Chiller and allow time for temperature stabilization
6) Prepare Plate for deep cleaning (PBS + 2% Tween 20 in 96 well plate using wells A1, B1, C1, D1)
7) Perform deep cleaning utility (perform steps 8,9) Once deep cleaning command reaches the 96 well plate perform steps 10,11,12,13)
8) Clean and fill Buffer Trough (rinse in dH20) Fill with 14 ml 1X sterile PBS
9) Clean and fill Cleaning Trough (rinse in dH20 from tap) Fill with 14 ml 1X sterile PBS + 1% Tween.
10) Fill Wash reservoir with dH20 from tap
11) Empty Waste reservoir (sink)
12) Fill enclosure humidification reservoir with dH20. Turn on enclosure pump.
13) Prepare source plate for printing. (Centrifuge source plate for 5 minutes @ 1100g to ensure that all air bubbles have been displaced.
14) Perform prime utility
15) Perform Tip Tuning (tune the tips to your samples for optimal performance. Use the buffer only it is an adequate representation of the protein solution)
Checklist prior to running array
1) Buffer trough covers must not be tilted
2) Glass slides well positioned
3) 96 well plate in correct position
4) No obstacles on or around stage
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