Bx60 system microscope

After mice were euthanized a tissue package containing prostate lobes, seminal vesicles and prostatic urethra was surgically excised and incubated in Tissue-Tek optimal cutting temperature compound (Sakura Finetek USA, Inc) on ice for 45 min, and then snap-frozen on dry ice in a cryomold. Sets of contiguous 15 µm thick tissue sections were cut using a CM1950 cryostat microtome (Leica Microsystems Inc) and arrayed onto SuperfrostPlus glass microscope slides (Thermo Scientific). The slides were exposed for 144 h on phosphor plates and read by a Fujifilm BAS-1800II bio-imaging analyzer (Fuji Photo Film Co.) generating digital images with 50 μm pixel dimensions. Digital images were obtained with an Olympus BX60 System Microscope (Olympus America, Inc.) equipped with a motorized stage (Prior Scientific, Inc.).

Single eyes from 21 subjects with retinopathy-positive CM included in the autopsy component of the research program (1996–2010) were analyzed as described previously [14 (link)]. We scored the histopathological severity of retinopathy according to a scale of intensity of retinal sequestration and maturation of sequestered parasites [14 (link)].
After fixation in 10% v/v buffered formalin, ocular specimens were opened horizontally in the pupil-optic nerve plane or by an equatorial incision. Gross pathology assessment was performed with a dissecting microscope, and retinopathy features were photographed. All samples were dehydrated and embedded in paraffin wax before 4-μm thick sections were cut with a manual rotary microtome (at least 100 sections per specimen). We investigated white-centered hemorrhages by making serial sections on 42 hemorrhages from 15 cases. Four hemorrhages were isolated by punch biopsy and embedded separately. Sequential sections were stained with hematoxylin and eosin (for staging and hemozoin), Martius-Scarlet-Blue (for fibrin), periodic acid-Schiff (for platelet-fibrin clots), and by immunohistochemistry to assess vessel integrity, clotting, and inflammation (Supplementary Table 1). We examined a minimum of 50 capillaries and 50 venules (diameter of 5–50 µm) per case using an Olympus BX60 system microscope.