Apparatus for high-throughput pollen extraction and counting

Abstract

An apparatus and method of quantitatively obtaining a measurement of pollen of a plant. One method of counting comprises imaging the sample with the pollen well-distributed in the focal plane of the imager. Image evaluation software can identify and count objects in the image that are consistent with pollen. Total pollen count for the plant can be derived from the count of pollen of the sample, proportionality of the sample volume to the starting volume, and proportionality of area of sample imaged to total area of sample. Pollen quantification can be used for research or commercial production decisions relative to the plant or its seed.

What is claimed is: 
     
         1 . An apparatus for counting pollen or similarly sized particles in a liquid having a characteristic that the particles tend to sink by gravity in the liquid, where the liquid and particles are contained in a sample container having a bottom generally in a plane, the apparatus comprising:
 a. a microscope having a field of view;   b. a stage positioned in the field of view of the microscope, the stage having a surface adapted to hold at least a portion of the bottom of the sample container in the microscope field of view; and   c. a digital camera operatively installed to image at least a portion of the field of view of the microscope at a focal plane that is at or near the surface of the stage,   wherein the digital camera is operatively installed so that a magnified digital image of a known portion of the bottom of the sample container, substantially focused at the plane of the bottom of the sample container, can be obtained.   
     
     
         2 . The apparatus of  claim 1 , further comprising a computer, wherein the computer is configured to store the magnified digital image. 
     
     
         3 . The apparatus of  claim 2 , wherein the computer further comprises image evaluation software adapted to count objects in the magnified digital image which appear to be a particle, and store a count of particles in the image. 
     
     
         4 . The apparatus of  claim 3 , wherein the computer is configured to correlate the stored count to identifying information about the particles or their origin. 
     
     
         5 . The apparatus of  claim 1 , wherein the particles are pollen of a plant. 
     
     
         6 . The apparatus of  claim 5 , wherein the pollen is pre-shed pollen. 
     
     
         7 . The apparatus of  claim 5 , wherein the pollen is post-shed pollen. 
     
     
         8 . The apparatus of  claim 1 , wherein the container is a well of a multiple well tray. 
     
     
         9 . The apparatus of  claim 8 , further comprising an actuator to move the stage relative to the microscope so that each well of the multiple well tray can be placed in the field of view of the microscope. 
     
     
         10 . The apparatus of  claim 9 , further comprising a computer, wherein a magnified digital image of each of two or more wells of the multiple well tray are stored by the computer. 
     
     
         11 . The apparatus of  claim 1 , further comprising:
 a. a blender comprising
 a blender container having an internal volume defined by a bottom, a top, and a sidewall radially disposed around a longitudinal axis and extending between the bottom and the top; 
 a motor in a base, and 
 a blade positioned in the internal volume at or near the bottom of the blender container; 
   b. a fluid pathway through the sidewall and intermediate of the top and bottom of the blender container, and in fluid communication with a sample extraction port;   c. a filter material positioned across the fluid pathway, the filter material having a filtering characteristic designed to allow passage of the particles; and   d. a valve mounted relative the fluid pathway and having a first position blocking fluid communication with the outlet port, and a second position unblocking fluid communication with the outlet port,   wherein the apparatus is configured to selectively extract a sample of a blended liquid and particle matrix through the outlet port when the valve is in the second position.