Standard Operating Procedure
Coulter Counter: 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
Date approved: 26 Jan 04
Procedure Overview: This procedure is used for quantifying corn pollen production per
tassel.
Equipment and reagents necessary:
Coulter counter
Isoton II Diluent (Beckman Coulter)
150 mL glass beakers
Pollen samples solutions (see SOP for pollen quantification)
1000 m Micropipette
Procedure:
1. Dilute pollen samples solutions before quantification in a 150 ml glass beaker
(typically, 1ml pollen sample solution in 50ml of isotone solution). Make sure to
shake the pollen sample solution until the dispersal of pollen grains is homogeneous
prior to pipetting out the 1 ml of sample. IMPORTANT: Pollen sample solution
volumes need to be measured before dumping.
2. The Coulter counter should be set to quantify particles between 50 and 90 m in
diameter (see SOP for Coulter counter operation). Set the dilution factor to reflect
appropriate total volume (in this case, 51 ml). Pollen quantity should be measured
using three consecutive readings. Save the outputs in the attached computer using
the Coulter counter software package.
3. After quantification, the pollen solution can be disposed of in the sink while flushing
with abundant quantities of water and the glass beakers washed with deionized
water before reuse.
4. Any remaining pollen sample solution can be stored in cold environment if further
quantifications are necessary. Otherwise solution can be poured down the sink drain
as mentioned above.
Personal Protective Equipment / Engineering Controls:
Safety glasses recommended
Gloves
Hazard controls:
None required
Waste Disposal Procedures & Decontamination:
Isoton II Diluent: Can be disposed of in the sink with abundant water.
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Health & Safety Info for required reagents:
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Isoton II Diluent
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Storage
precautions and
incompatibilities
No special precautions;
no hazardous
incompatibilities
None
determined
The above summary consists of guidelines for proper handling & disposal
of chemicals used in this procedure. You must read attached MSDSs for
more specific information before using the procedure.
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