14
Stimulation Indices and Statistical Analysis
Means and standard deviations were determined for each data group.
An F-test was used for variance analysis. The Student-t test was used
to determine the statistical significance of increases over control
values (20,103). A 95% or greater confidence level (p <_ 0.05) was used
for both the F-test and the t-test.
Stimulation indices were used to express increases of mitogen
stimulated cultures over control cultures or mixed lymphocyte cultures
(MLC's) over controls. Indices for mitogenic studies were determined
using the following equation: Mean CPM of stimulated cultures Indices
Mean CPM of control cultures
for MLC studies were calculated by using the following formula:
Mean CPM of MLC between Fish A and Fish B
(Mean CPM of Fish A Control Culture + Mean CPM of Fish B Control) t 2
Histological and Morphological Techniques
Serial cross sections of paraffin embedded gill regions of bluegill
were kindly prepared by Mr. Melvin Laite (Department of Pathology, J.
Hillis Miller Health Center, Gainesville, Fla.). Sectioned tissues were
*
stained with hematoxylin and eosin.
Cell suspension smears or cytocentrifuge (Shandon-Elliot Inc.)
preparations were stained with May-Grunwald-Giemsa for morphological
examination.
Autoradiography
*7
Cultured cells, pulsed with H-thymidine for 24 hr, were pipeted
from microculture tray wells, washed three times with RPMI 1640 and
cytocentrifuged. Cytocentrifuged preparations were coated with nuclear
track emulsion (Kodak, Inc., Rochester, N.Y.; type NTB3), exposed,