z2coultercounteroperationfor.docVIP

Z2 Coulter Counter operation for Quantifying corn pollen production Department: Agronomy Created by: Agustin Fonseca and Sebastian Schneider Laboratory: Crop Production Physiology Supervisor: Mark Westgate Lab Supervisor: Maria Hartt Eckerman Overview The Coulter counter provides the ability to count particles (in this case, pollen grains) relative to user-defined thresholds. This protocol briefly describes the start up procedure, calibration of the machine, operation and sample analysis, and shutdown procedures. For more details, the Beckman Coulter Z series manual is located in a drawer underneath the machine (page references are provided in this guide). Start up procedures Before turning on the Coulter counter be sure that the waste jar is empty and that the diluent jar is full with Isoton II diluent (13). Waste Isoton II diluent can be disposed of in the sink with abundant water. Pollen counting requires a 200 (m aperture tube. If a different aperture is in place, replace it with the 200 (m (pages 2-15 and 4-19). Currently this is the only aperture we have, so this step is not required. Turn on the Coulter counter. Calibration (1) Beckman recommends that the user check the size calibration for each aperture tube at least once a month to monitor for any system variations. However, when a lot of samples are being analyzed, we recommend doing so more frequently. Prepare a calibration suspension by adding 5 drops of the 10 (m standard into a beaker containing 50 ml of Isoton II diluent. Place the beaker of the calibration suspension on the platform and raise it until the aperture tube, electrode and stirrer are immersed. Turn on stirrer (knob on top front of Coulter counter); adjust speed to a gentle but thorough stir (stirrer will automatically stop when platform is lowered). Press Set-up and screen S1 will display. At Selected units select (m by using the cursor keys. Press Set-up and screen S2 displays. At Select Apert