Dfc450 c digital camera

To examine the effect of dietary hempseed on the density of bone marrow adipocytes, immunohistochemistry staining for the marker PRDM16 protein was employed. This zinc finger transcription factor plays an important role in the differentiation of adipocytes within the bone marrow [45 (link),46 (link),47 (link)]. Tissue was stained using a rabbit anti-human polyclonal antibody targeting PRDM16 (ThermoFisher Scientific, Waltham, MA, USA; catalog #PA5-20872). All other reagents were obtained from the Pierce Peroxidase Detection Kit (ThermoFisher Scientific; catalog #36000) and the protocol was followed according to the manufacturer’s instructions. Briefly, a goat anti-rabbit secondary antibody was applied to the tissues, followed by an anti-goat strep-HRP tertiary antibody. Samples were counterstained with hematoxylin, dehydrated and mounted. Slides were analyzed for PRDM16-positive cells (staining intensity) using a Leica DM6B widefield microscope and imaged with the attached Leica DFC450-C digital camera. Quantitative values were obtained by counting PRDM16-positive cells (brown stain) and total cells (blue stain) in the bone marrow and calculating a percentage.

For histopathological and morphometric analyses, the lungs were perfused with 5 mL of PBS containing 10% antibiotics (Penicillin-Streptomycin—Cat# 15140148, Gibco™, Waltham, MA USA) through the right ventricle of the heart. A fragment of the left lung was excised and immediately placed in the methacarn solution for fixation. After fixation, the lung fragments were dehydrated, clarified in xylol, embedded in paraffin, and sectioned. Thereafter, 5 µm-thick sections were prepared by slicing the tissue at 135 µm intervals. The sections were stained with hematoxylin and eosin (H&E), Grocott–Gomori methenamine silver (GMS), or mucicarmine (specific for Cryptococcus spp.) and evaluated using light microscopy (LMD6 system and motorized stage; Leica, Wetzlar, Germany; XYZ module LX; objectives HC PL FL 1.25×/0.04, 10×/0.30, 20×/0.40, 40×/0.60) and a DFC450 C digital camera (Leica). The images were processed using ImageJ Fiji open-source image processor software. Quantification of the number of yeast cells, average yeast area, yeast frequency, and compromised pulmonary parenchyma was performed in a semi-automated quantitative manner using ImageJ Fiji.

Colon cancer cell lines were grown in 6-well plates. Wounds were made in a confluent monolayer of each well by using a 10 μL pipette tip. The cells were then treated with 50 μg/mL of the methanolic, acetonic, or aqueous extract and the wound closure was compared with that of untreated cells at 0, 6, and 24 h after wounding [27 (link)]. Digital photographs were captured with a LEICA DFC450 C digital camera (LEICA Microsystems, DE). The percentage of wound closure was calculated through the comparison of the wound areas before and after stimulation using the following formula:
Wound closure (%) = (initial scratch size – size of the scratch after an identified culture period) / (initial scratch size) × 100.

The images of Petri dishes and protonemal tissues including gametophores were photographed using a digital camera (EOS6D; Canon, Tokyo, Japan), a microscope over-eyepiece camera, or tereomicroscopes (MZ10F; Leica Microsystems, Wetzlar, Germany) fitted with a DFC450 C Digital Camera. The magnified images of gametangia and sporophytes were observed by inverted microscope (Axio Observer D1, Zeiss).