**4. Histological studies on the liver, kidney and pancreas**

The effect of the extracts on tissue architecture of the pancreas of treated diabetic rats was evaluated by histological studies of tissue sections obtained from the animals. On day 28 of the experiment, one animal was randomly selected from the different groups and sacrificed by over-dose of chloroform anaesthesia. The whole pancreas from each animal was removed and placed in 10% formalin in normal saline for histological studies. The isolated organ was placed in an automatic tissue processor for 24 hrs. After 24 hrs, the tissues were solidified in molten wax and sectioned using automatic tissue sectioner. The tissue sections were then fixed on slides with haematoxylin and eosin. The stained slides were fixed with mountant, allowed to dry and viewed under the microscope (x400). This procedure was repeated for the liver and kidney collected from the sacrificed animals.

#### **4.1. Statistical analysis**

counted beginning at the left of the top row of small squares, then from right to left for the

Cells counted x 10(0.1mm depth) x 5(1/5 of sqmm) x 200(1:200) dilution = erythrocytes per

The sum of the cells in the five small squares multiplied by 10,000 = total erythrocytes per cu

Leucocyte diluting pipetting was used to draw the blood sample to a point marked 0.5 and filled with leucocyte diluting fluid up to the 11mark above the bulb. The mixture was shak‐ en for 3minutes until well mixed. Two to three drops from the pipette was discarded before filling the counting chamber of haemocytometer. The leucocytes were allowed to settle for 1minute. The leucocytes in the larger squares of haemocytometer chamber were counted

The sum of the cell counted in the 4 corner squares multiplied by 50 = total leucocytes per

The effect of the extracts on tissue architecture of the pancreas of treated diabetic rats was evaluated by histological studies of tissue sections obtained from the animals. On day 28 of the experiment, one animal was randomly selected from the different groups and sacrificed by over-dose of chloroform anaesthesia. The whole pancreas from each animal was removed and placed in 10% formalin in normal saline for histological studies. The isolated organ was placed in an automatic tissue processor for 24 hrs. After 24 hrs, the tissues were solidified in molten wax and sectioned using automatic tissue sectioner. The tissue sections were then fixed on slides with haematoxylin and eosin. The stained slides were fixed with mountant, allowed to dry and viewed under the microscope (x400). This procedure was repeated for

and multiplied by 50 to obtain the total number of white blood cells (Coles, 1986).

Cells counted x 20 (1:20 dilution) x 10(0.1mm depth)/4 (no of sq mm counted)

**4. Histological studies on the liver, kidney and pancreas**

the liver and kidney collected from the sacrificed animals.

next row and so on.

120 Antioxidant-Antidiabetic Agents and Human Health

mm (Coles, 1986)

Calculation:

OR

cubic mm.

= WBC/cubic mm

**3.9. Determination of leucocyte (wbc) count**

Calculation

cu mm.

OR

Data was analyzed using graph pad prism 5 and the results expressed as mean ± SD. The results were further subjected to one way ANOVA for comparisons and differences between means were considered significant at P< 0.05.
