Dfc420 camera

The thin-sections have been prepared in the laboratory of the Institute of Geosciences at the Rheinische Friedrich-Wilhelms-Universität Bonn, Germany. All bones were sectioned according to Stein & Sander (2009) and Lamm (2013) , however, their technique was slightly modified. Wet silicon carbide (SiC) grinding powder with grit sizes of 600 and 800 were used for grinding and polishing the thin-sections. All limb bone sections were cut at the midshaft plane. A sagittal section of the intercentrum was made. The osteohistological analysis was performed with a LEICA DM LP polarizing light microscope and the photographs were taken with a LEICA DFC 420 camera attached to the microscope. The sections were scanned with an EPSON PERFECTION 750V PRO scanner in order to gain microstructural overview.

Immunohistochemistry from the carotid arteries was performed on formalin-fixed, paraffin-embedded tissue sections. 4 μm slides were then deparaffinated using xylol and ethanol.
Samples were incubated with the anti-CSE primary monoclonal antibody (12217-1-AP Proteintech, Chicago, IL, USA) at a dilution of 1 : 200. Other slides of the same samples were incubated with antihemoglobin (clone: goat polyclonal HRP (ab19362), Proteintech Group, Rosemont, IL 60018, USA) primary monoclonal antibody at a dilution of 1 : 100; with HO-1 (clone: rabbit polyclonal (10701-1-AP), Proteintech Group, Rosemont, IL 60018, USA) primary monoclonal antibody at a dilution of 1 : 200; and with HO-2 (clone: rabbit polyclonal (14817-1-AP), Proteintech Group, Rosemont, IL 60018, USA) primary monoclonal antibody at a dilution of 1 : 400. Specific antibody binding was visualized by the Dako EnVision FLEX/HRP and FLEX DAB3 Chromogen detection system (Dako, Glostrup, Danmark) followed by hematoxylin counterstaining and coverage. The intensity and distribution of protein immunostaining were assessed by light microscopy (Leica DM2500 microscope, DFC 420 camera and Leica Application Suite V3 software, Leica).

Twenty seeds per replication were randomly selected from each inoculation time point of each cultivar for the MoT sporulation assay. Three replicates were prepared for each treatment. Surface sterilized seeds were placed in sterile micro-slides (76 × 26 mm, Glaswarenfabrik Karl Hecht GmbH and Co KG, Germany) in a 90 mm Petri dish (single seed placed in single slide) containing sterilized distilled water-soaked sterile filter paper to maintain humidity. Twenty-four to 48 h after incubation, seeds were observed under a stereomicroscope (Leica Wild M10) followed by a compound light microscope (Leica Leitz DM RB). The samples were photographed with a Leica DFC420 camera (version 2.8.0.0), and all images were processed using Leica Application Suite V4 (LAS V4) software (version 4.9.0.129). Subsequently, a single infected seed presenting MoT conidia was placed in a 1.5 ml Eppendorf tube containing 1 ml of SDW and vortexed briefly. After vortexing, 5 μl of suspension was poured into each counting chamber of hemocytometer (Fuchs-Rosenthal, 0.0625 mm²) to check the homogeneity of the conidial suspension before the conidia were counted. This procedure was repeated five times, and the average counts were multiplied by 200 to obtain the total number of conidia per seed.

Anthers were fixed in FAA (10% formaldehyde, 3% acetic acid and 43.5% ethanol) placed under vacuum for 1 h and then keep room temperature. After dehydration in a graded ethanol series and diaphaneity in clearing medium, the material was embedded in Paraffins (from HuaShenPai). Sections (6 μm) were obtained with a Leica Reichert Supernova microtome, placed on glass slides, and stained with a solution of 1%(w/v) toluidine blue O (toluidine blue O 1 g, 95% alcohol 4 mL, 10% acetic acid 10 mL, distilled water 86 mL). Sections examined using a Leica fluorescent compound microscope and Images were captured with a Leica DFC420 camera, and processed with Leica Application Suite software.
The images of whole morphology of WT and mutant were captured using SONY DSC-H50 camera. And the flowers, siliques and seeds morphology were examined using a Leica dissecting microscope.

Spleens were removed from 1 year old wild type or RetSat^−/− mice and fixed in formaldehyde (4% in phosphate buffer) for 1 day. Dehydration was carried out before embedding into paraffin according to standard protocol. After paraffin became solid, blocks were cut with a microtome to obtain 3–4 micrometer thick sections. After the deparaffination and hematoxylin and eosin (HE) staining, immunohistochemical staining was performed using anti-Cleaved Caspase-3 (Cell Signaling Technology, Inc, Global Headquarters USA) primary monoclonal antibody at a dilution of 1:300 and an horse radish peroxidase (HRP)-labelled polymer anti-rabbit secondary antibody (Dako, Glostrup, Denmark). The intensity and distribution of immunostaining was assessed by light microscopy (Leica DM2500 microscope, DFC 420 camera and Leica Application Suite V3 software; Leica).

Chondrocytes were cultured and administrated with 1 μM PGE₂ and TMF. After 48 h treatment, cells were harvested. 4% paraformaldehyde was employed to fix the cells. A tunnel and DAPI staining kit (Abcam, USA) was used for determining the cell apoptosis. A Leica DM3000 microscope was employed for all fluorescent images. And DFC 420 camera (Leica, Germany) functions to take the photos.