blood cells lab 18

All avian and reptile blood cells are nucleated, therefore, leukocyte counts must be done manually with a hemacytometer. The purpose of this lab was to accurately determine the red blood cell count of one rainbow trout and one goose with a hemacytometer. A Hemacytometer is a specialized microscope slide with two grids, of known dimensions, etched on the central surface.

Élie Metchnikoff (1845-1916)
Russian biologist who introduced the theory of phagocytosis; that certain white blood cells are able to engulf and destroy harmful substances such as bacteria. This lead to modern immunology.

Procedure

Red Blood Cell Count

Dr. Allen made 200 fold dilutions of the blood from each fish and the goose for us. This was done to enable us to differentiate the various cell types. We first placed a cover slip over the grid on the hemocytomerer slide. A small sample of diluted blood was then carefully applied to the edge of the cover slip using a micropipeter. The blood should flow under the cover slip and over the grid by capillary action. However, you have to be careful not to apply too much. You don’t want any of the dilute blood sample to overflow down into the moat. Using the 10X objective on the microscope, we then counted the total number of red blood cells that are in the five numbered squares. The red blood cells that fell on the line of the grid were only counted if they were on top of, or to the right side of the line of the square.

Equipment

	The hemocytometer grid
Squares 1-5 are used for red blood cell counts and squares A-D are used for white blood cells.

Results
To get the cell count in number of cells/µl, we multiplied the total number of cells in the five squares by 10,000.

My group did trout #3:	For square 1 we got a total of 25 RBC X 10,000 = 250,000
	For square 2 we got a total of 36 RBC X 10,000 = 360,000
	For square 3 we got a total of 32 RBC X 10,000 = 320,000
	For square 4 we got a total of 27 RBC X 10,000 = 270,000
	For square 5 we got a total of 34 RBC X 10,000 = 340,000
	Total =1,540,000

My group also did RBC counts of the female chicken:	For square 1 we got a total of 44 RBC X 10,000 = 440,000
	For square 2 we got a total of 40 RBC X 10,000 = 400,000
	For square 3 we got a total of 42 RBC X 10,000 = 420,000
	For square 4 we got a total of 43 RBC X 10,000 = 430,000
	For square 5 we got a total of 45 RBC X 10,000 = 450,000
	Total =2,140,000

ZOOLOGY 310 GUIDE TO BLOOD CELLS