Intraglomerular deposition of low-density lipoprotein (LDL) and oxidized LDL has been described in various human glomerular diseases. Yet it is not clear whether esterified cholesterol (EC) and unesterified cholesterol (UC) carried in LDL are mobilized from deposited LDL particles or accumulate in the diseased human glomeruli, particularly in the segmentally sclerotic lesions. To address this issue, frozen sections of renal biopsies were first immunostained to localize apolipoprotein B (apo B) and then oil red O (ORO) stained to colocalize neutral lipids. By using 124 ORO-positive biopsies and nine ORO-negative ones, UC was visualized directly with filipin staining, and EC was visualized after its enzymatic hydrolysis and staining with filipin. Seventy-seven biopsies (58%) showed filipin staining of accumulated EC and/or UC in the glomeruli. Of these, 11 showed heavy filipin staining for both EC and UC in the segmentally sclerotic lesions. In a group with UC deposits in the sclerotic segments, the percentage of the glomeruli affected by sclerosis and the intensity of filipin fluorescence for UC were significantly higher than biopsies with only mesangial UC deposits. Most filipin-positive biopsies showed apo B staining mainly in the mesangium. Yet in the sclerotic segments, apo B staining was rarely noted. Accumulated apo B-stained lipoprotein was not coincident with ORO-stained lipid in the diseased glomeruli. These results suggest that both EC and UC accumulate in the sclerotic glomerular segments as the glomerular lesions are advanced, and that these EC and UC appear to be derived from altered LDL with progressive loss of apo B.