Dp71 camera

Intracellular ROS production was measured using DCFH-DA as described previously [55 (link)]. Briefly, HepG2 cells were grown to confluence in 6-well plates and treated with ZEN for 2 h. Following treatment, cells were incubated with a final concentration of 10 μM DCFH-DA for 30 min. After two rounds of washing with cold PBS, the production of ROS (H₂O₂) was visualized using an Olympus IX71 fluorescence microscope and images were digitally captured with an Olympus DP71 camera and DP controller software (Olympus Optical Co., Ltd, Tokyo, Japan). To measure the production of intracellular superoxide anion, HepG2 cells were treated with ZEN for 2 h. The cells were then incubated with a fluorogenic dye, DHE (1 μM, 30 min), followed by cold PBS washes. The cells on coverslips were fixed with 4% formaldehyde and visualized with a super-resolution confocal laser scanning microscope (Carl Zeiss Co., Ltd, Oberkochen, Germany). Quantification of DCFH-DA positive area (green fluorescence) and DHE positive area (red fluorescence) was conducted using Image J software (National Institute of Health, Bethesda, MD, USA) and graphically presented.

To confirm the development of typical lipid-rich atherosclerotic plaques, the excised aortas were imaged intact. They then underwent Oil-Red-O staining. All photographic images were obtained by a digital camera.
To evaluate noninvasive imaging of atherosclerotic lesions, histopathology was performed on in vivo imaged aortas with an intense ^99mTc-IDA-D-[c(RGDfK)]₂ signal. After undergoing SPECT/CT and autoradiography imaging, the excised aortas were fixed with 10% formalin, embedded in paraffin, cut into 5-μm sections and deparaffinized. The sections were subsequently stained with hematoxylin and eosin or Masson’s trichrome stain to characterize the morphology and composition of the recorded peak signal of the aorta. Bright field color micrographs were obtained on a BX51 microscope equipped with DP71 camera (Olympus Optical Co., Ltd., Tokyo, Japan).

The humeri of the euthanized animals were carefully removed, cleaned, and measured for length with a digital caliper. They were then fixed in 4% neutral buffered formalin for 48 h at room temperature, decalcified with Surgipath Decalcifier II (Leica Biosystems Richmond, Inc. USA) for several hours (depending upon the age of the animal), dehydrated through graded ethanol series (70, 95, and 100%), and stabilized by two sequential changes of chloroform for paraffin embedding. Histological studies and EGP height measurements were performed on deparaffinized sections of 6 μm thickness that had been stained with hematoxylin-eosin and Alcian blue. The height of the EGP was measured by drawing a straight line from the apical border of the reserve zone cells to the lower border of the mineralized cartilage. The findings presented here represent the average of at least five measurements per each section. The slides were photographed under an Olympus BX40 microscope equipped with an Olympus DP71 camera (Olympus Optical Co. GmbH, Hamburg, Germany), and analyzed with Image-Pro software (version 4.5.1.22, Media Cybernetics, Inc., Rockville, MD, USA).

MCF-7 cells were grown to confluence in 6-well plates. Cells were pre-treated with or without 5 mM NAC for 1 h followed by PL treatment (0, 5, 10, and 20 µM) for 3 h. Following the treatments, cells were incubated with 2’,7’-dichlorofluorescin diacetate (DCFH-DA) (final concentration, 20 μM) at 37 °C in a 5% CO₂ incubator for 30 min. Cells were washed 3× with PBS to terminate the reaction. The generation of H₂O₂ was evaluated using an Olympus IX71 fluorescence microscope (Olympus Optical Co. Ltd., Tokyo, Japan) and the fluorescent images were captured using an Olympus DP71 camera and DP controller software, version 2.2 (Olympus Optical Co. Ltd., Tokyo, Japan).

The humeri were fixed in 10% neutral buffered formalin for 48 h at room temperature, decalcified with Surgipath Decalcifier II (Leica Biosystems Richmond, Inc., Richmond, IL, USA) for several hours (depending on the age of the animal), dehydrated through a graded ethanol series (70%, 95%, and 100%), and stabilized by 2 sequential changes in chloroform for paraffin embedding. Histological studies and EGP height measurements were performed on deparaffinized 6 μm thick sections stained with hematoxylin-eosin and Alcian blue. For each bone, two sections were measured, for a total of five measurements per section. The height of the EGP was measured by drawing a straight line from the apical border of the reserve zone cells to the lower border of the mineralized cartilage. The proliferative and hypertrophic zones were measured separately from the apical border to the first hypertrophic cell and then from there to the lower border to the EGP. The slides were photographed under an Olympus BX40 microscope equipped with an Olympus DP71 camera (Olympus Optical Co. GmbH, Hamburg, Germany) and analyzed with Image-Pro software (version 4.5.1.22, Media Cybernetics, Inc., Rockville, MD, USA) by two specialists blind to their origin. The findings below represent the average of at least five measurements for each section.