Retina cross-sections were mounted and stained with toluidine blue. Images of whole retina cross-sections were obtained using Leica leitz Aristoplan microscope equipped with DFC 480 camera (Leica) and LAS 3.7 version software. Thickness of the different nuclear and synaptic layers were determined using Visilog 6.4 version software (Noesis).
Dfc480 camera
Manufactured by Leica
Sourced in Germany, United States
The DFC480 is a camera designed for use in laboratory environments. It features a high-resolution sensor and advanced image capture capabilities to facilitate detailed documentation and analysis of microscopic samples.
Automatically generated - may contain errors
Lab products found in correlation
Las v 4, by Leica (3 mentions)
Dm4500b microscope, by Leica (2 mentions)
Live dead baclight bacterial viability kit reagents, by Thermo Fisher Scientific (2 mentions)
Fv1000 confocal inverted microscope, by Leica (2 mentions)
As lmd laser dissection microscope, by Leica (2 mentions)
Dm5500b microscope, by Leica (2 mentions)
Leitz aristoplan microscope, by Leica (1 mentions)
Fsc 22, by Leica (1 mentions)
Dmi6000, by Leica (1 mentions)
Anti β2, by Abcam (1 mentions)
Aquamount, by Merck Group (1 mentions)
Pt link, by Agilent Technologies (1 mentions)
Autostainer, by Agilent Technologies (1 mentions)
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
Dm2500 microscope, by Leica (1 mentions)
Dm lb2 microscope, by Leica (1 mentions)
Stemi 2000 c, by Zeiss (1 mentions)
M2 axioimager, by Zeiss (1 mentions)
Axiocam erc5s camera, by Zeiss (1 mentions)
Calcofluor white, by Merck Group (1 mentions)
25 protocols using dfc480 camera
1
Retinal Layer Thickness Analysis
2
Histological Analysis of Liver and Adipose Tissues
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For histologic examination, liver and adipose tissues of mice were embedded in frozen section media (FSC 22, Leica, Jena, Germany) and frozen at −20 °C (liver), or −25 °C (adipose tissue). Specimens were prepared by cutting samples into 8 μm thickness using CM 1860 cryomicrotome system (Leica, Jena, Germany). Specimen slides (Marienfeld, Lauda-Königshofen, Germany) was dried and fixed in 10% formalin solution for 5 min. Liver sections were stained with Oil Red O (dissolved in propylene glycol) for 30 min and Mayer’s Hematoxylin as counterstain for 1 min. Liver and adipose tissue were stained with Hematoxylin & Eosin to examine structural changes. Stained slides were examined under microscope (DMI 6000, Leica, Germany) equipped with DFC480 camera (Leica, Germany).
3
Immunohistochemical Analysis of Tumor Biomarkers
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The spheroids were fixed in Carnoy fixative (Ethanol:Chloroform:Glacial acetic acid 6:3:1) and embedded in paraffin. Patient tumour tissues were fixed in neutralized formalin and embedded in paraffin. Antigen retrieval was performed at low pH retrieval buffer (EnVision® Flex Target Retrieval Solution Low pH) at PT-Link (Dako). Immunohistochemical analysis was performed on an automated immunostainer (Dako Autostainer) using DakoCytomation EnVision®+ System-HRP (DAB) according to the manufacturer’s instructions. For CA IX staining, the sections were labelled with M75 antibody (hybridoma medium) diluted 1:100 for 1 h at room temperature. For β1 and β2 adrenoreceptors and Ki-67 the sections were incubated with anti β1 (Invitrogen, Carlsbad, CA, USA, 1:500), anti β2 (Abcam, 1:250) overnight at 4 °C, for Ki-67 (DAKO, 1:100) for 1h at room temperature and, after washing, incubated with secondary anti mouse-HRP or anti rabbit-HRP antibody for 30 min at room temperature. Staining was visualized using 3,3’-diaminobenzidine (DAB) as a chromogenic substrate for 1 min. The sections were counterstained with Mayer’s haematoxylin and mounted in Aquamount (Merck, Darmstadt, Germany). The stained sections were examined using a Leica DM4500B microscope and images were captured by a Leica DFC480 camera.
4
Cell Viability Imaging on Collagen I
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Cells were cultured on 0.2% collagen type I coated coverslip. NUCLEAR‐ID® Blue/Red cell viability reagent was added to the cells at dilution 1:1,000 for 30 min at 37°C. Then, the cells were fixed with Fluorescent Mounting Medium (Dako) on Superfrost Ultra Plus Microscope Slide (Thermo Scientific). Images were obtained with an immunofluorescence microscope (Leica DM2500 microscope, Leica DFC480 camera).
5
Fluorescence Microscopy Imaging Protocol
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Epifluorescence images were collected using the Leica DM LB 2 microscope equipped with the LAS acquisition software and a Leica DFC480 camera for detection. Confocal images were collected using a Zeiss (Oberkochen, Germany) Axiovert 200M inverted microscope equipped with the LSM 5 Pascal point laser module, the LSM AIM acquisition software, and 2 PMT detectors for spectral detection. Merging of images, analysis and quantification were performed with the Image J, Volocity, and Imaris softwares.
6
Microscopy Analysis of P. aeruginosa Biofilms
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For microscopy analysis, narrow glass coverslips (10.5 × 35 mm, ProSciTech, Townsville, Australia) were inserted in the biofilm culture tubes and P. aeruginosa biofilms were grown otherwise as described above. After 24 h incubation and 20 min treatment, the coverslips harbouring biofilms were carefully removed from the tubes by using sterile tweezers and rinsed twice with PBS before wiping clean one side and staining the other side with LIVE/DEAD BacLight bacterial viability kit reagents (Molecular Probes) according to the manufacturer’s procedure. Three hundred microliters of the staining solution (1:1,000 dilution of each SYTO9 and propidium iodide components in PBS) was trapped between the biofilm sample on the coverslip and a standard microscope slide. After 20 min incubation at room temperature in the dark, the samples were observed with an Olympus FV1000 Confocal Inverted Microscope, and imaged with Leica DFC 480 camera. Cells that were stained green were considered to be viable, those that stained red were considered to be dead.
7
Seed Coat Developmental Imaging Techniques
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Ruthenium red staining followed the protocol from Arsovski et al. (2009) with modifications excluding pre-hydration and imbibition for 10 min. Imaging was conducted using a Zeiss Stemi 2000-C dissecting microscope with an attached AxioCam ERc 5s camera. For calcofluor white staining, mature seeds were imbibed in a solution of 0.002% (w/v) calcofluor white (Sigma-Aldrich, F3543) with 0.01% Triton X-100 (Sigma-Aldrich, T8532) in 100 ml Tris (pH 8.0) for 10 min and imaged using a Leica AS LMD laser dissection microscope with an attached DFC 480 camera. To confirm developmental stages, seeds were cleared as per Tucker et al. (2012) (link) using Hoyer’s light solution (Anderson, 1954 (link)) and observed using differential interference contrast (DIC) and Nomarski optics on a Zeiss M2 Axio imager. To observe anatomical details of seed coat development, staged seed samples were fixed in 0.25% glutaraldehyde, 4% paraformaldehyde and 4% sucrose in phosphate-buffered saline (PBS; pH 7.2), embedded in LR-White resin and sectioned to 1.0 μm before staining in 0.01% toluidine blue in 0.1% sodium tetraborate as per Aditya et al. (2015) (link).
8
Immunohistochemical Detection of CAIX
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Paraffin-embedded tissues were sectioned and mounted onto glass slides. The 5 μm-thick slides were deparaffinized and rehydrated. CAIX was detected using the DAKO EnVision™ FLEX System according to the manufacturer’s instructions. Sections were incubated with anti-CAIX antibody M75 (1:25) over night at 4 °C. Negative controls were prepared by omission of the primary antibody. The stained sections were examined using a Leica DM4500B microscope and images were captured with a Leica DFC480 camera.
9
Zebrafish Model of PNPLA6 Mutations
10
Quantification of Intestinal Immune Cells
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Paraformaldehyde-fixed sections were paraffin embedded, sectioned and stained with haematoxylin and eosin for enumeration of eosinophils and villus: crypt ratios. An additional section from the jejunum was processed for immunohistochemical determination of T-cells, macrophages and Foxp3+ cells using the follow antibodies, respectively: rabbit anti-human CD3 (Dako), mouse anti-human macrophage (clone MAC387, Abcam) and mouse anti-human Foxp3 (clone 22510, Abcam). Appropriate isotype controls were used to determine antibody specificity. Bound antibodies were visualised with the Ultravision LP Detection System and aminoethyl carbazole substrate (ThermoFisher Scientific, Denmark). The Carnoys-fixed jejunum section was stained with Toluidine blue for mast cell enumeration. For determination of cell numbers, 10 random fields of each tissue section were counted by a single blinded observer using a calibrated counting grid covering a total area of 0.25mm2 at 400x magnification. Cell counts were expressed as cells/mm2 tissue. For villous: crypt ratios, 10 well-orientated villus: crypt units were randomly selected from each tissue section by a blinded observer. The sections were photographed using a Leica DFC480 camera and measurements performed using LAS v4.6 software (Leica, Switzerland).
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