RBC counts in Turkeys
Evenson, Don Chem (evensond@mg.sdstate.edu)
Tue, 24 Jan 95 14:29:00 cst
In response to David Abbott:
We often use chicken blood as a DNA content
marker for other cell types and would expect turkey blood cells to behave in
a similar fashion. Using the two-step AO techniques developed by
Darzynkiewicz and colleagues the RBCs have a tight cluster (green vs red
fluorescence) and the other blood cells have a significantly higher red
fluorescence and thus are easily resolved; the CV is in the range of 2.5
using frozen cells. For absolute count of cells we use a method
originally described by Steinkamp and Crissman where a known concentration of
fluorescent latex beads are mixed with the cells of unknown concentration.
This mixture of beads and cells are measured (green vs red fluorescence),
gated and calculations are made to determine the concentration of the cell
type on a per ml basis. Several factors that need careful attention paid to
are: 1) very careful counting (hemocytometer and/ or coulter counter) of the
bead concentration, 2) vortex the bead stock before each mixing with the cell
type, and 3) accurate pipetting. Using this method we recently published a
paper "Rapid Determination on Sperm Cell Concentration in Bovine Semen by
Flow Cytometry" Evenson et al, J. Dairy Sci., 76:86-94. Some bull semen
cooperatives use this technique to determine the absolute count of sperm
which is of great economic interest to these businesses. We concluded in
that paper that this method is the most accurate and precise for
determination of absolute sperm count. The same principles should work well
for counting your turkey RBCs.
Don Evenson, Ph.D.
Olson Biochemistry Labs
South Dakota State University
ASC 136, Box 2170
Brookings, SD 57007
phone: 605-688-5474
FAX: 605-688-6295
Email: evensond@mg.sdstate.edu
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