Figure 8
Neutral lipid accumulation in PHA-stimulated PBMCs from control 2 and AD-FDR subjects. Immediately after separation cells were incubated with PHA for the indicated time periods. After harvesting, cells were washed, fixed by soaking in 10% formalin, stained with ORO for NL, and counter-stained with Mayer's hematoxylin for nuclei. Cells were then examined by light microscopy and two different fields per sample were imaged. Red ORO intensity was measured in these two fields by using NIH Image J software. Panel A shows images of ORO staining of one control 2 (47 years old) and two AD-FDR one (48 years old) with ORO score 0 and the other one (37 years old) with ORO score 3 at 0 time. These images are representative of at least 30 control 2, 15 AD-FDR with ORO score between 0 and 1, and 15 with ORO score between 2 and 3. Panel B shows red intensity expressed as mean pixels ± SE/cm2 of triplicate wells obtained by utilizing four created different regions of interest (ROIs). *P < 0.01, compared with control 2 by Student's t test.