The EnVision detection system HRP/DAB+ (Dako) was used as previously described (Kim et al., 2009 (link)). Briefly, tissue sections were deparaffinized in xylene and rehydrated in ethanol following treatment in pre-heated target retrieval solution. Following washes, serum-free blocking solution was applied for 40 min at RT. In-house anti-HDAC9 monoclonal antibody was used overnight at 4°C then treated with polymer/HRP and DAB. After washes, the slides were counterstained with hematoxylin, dried and mounted with Permount. Photomicrographs were captured using an Olympus BX41 dual head light microscope equipped with an Olympus Q-Color 5 digital camera (Olympus America).
Q color 5 digital camera
Manufactured by Olympus
Sourced in Japan
The Q-Color 5 is a digital camera designed for laboratory use. It captures high-quality images and provides accurate color representation for scientific and medical applications. The camera features a high-resolution sensor and advanced image processing capabilities to deliver consistent and reliable results.
Automatically generated - may contain errors
Lab products found in correlation
Bx51 microscope, by Olympus (5 mentions)
Bx41 microscope, by Olympus (4 mentions)
Q capture suite acquisition software, by Olympus (3 mentions)
Oct compound, by Sakura Finetek (2 mentions)
Bx41 dual head light microscope, by Olympus (1 mentions)
Reactive oxygen species assay kit, by Beyotime (1 mentions)
Thioflavine s, by Merck Group (1 mentions)
Trueblack lipofuscin autofluorescence quencher, by Biotium (1 mentions)
Everbrite mounting media, by Biotium (1 mentions)
Four chamber slide, by Thermo Fisher Scientific (1 mentions)
Bx53 compound microscope, by Olympus (1 mentions)
Sf93 20, by Thermo Fisher Scientific (1 mentions)
Millipore membrane, by Merck Group (1 mentions)
Alexa fluor 568 secondary antibody, by Thermo Fisher Scientific (1 mentions)
3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions)
Alexa fluor 488 secondary antibody, by Thermo Fisher Scientific (1 mentions)
Mom peroxidase kit, by Vector Laboratories (1 mentions)
Vectashield mounting media with dapi, by Vector Laboratories (1 mentions)
Prolong gold antifade solution, by Thermo Fisher Scientific (1 mentions)
Alexa 488, by Merck Group (1 mentions)
17 protocols using q color 5 digital camera
1
Immunohistochemical Detection of HDAC9
2
Oxidative Stress and Hepatocyte Apoptosis
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The level of intracellular ROS was quantified using the Reactive Oxygen Species Assay Kit (Beyotime, China). Primary hepatocytes were obtained as described [1 (link)], and the cells were washed three times with PBS. DCFH-DA (10 μM) was added, and the cells were incubated for 30 minutes at 37°C in the dark. After the cells were washed three times with PBS, photomicrographs were obtained using an Olympus BX41 microscope (Center Valley, PA) equipped with an Olympus Q-color5 digital camera. The CAT, MDA, GSH, and SOD activities of the liver tissue were measured using commercial kits according to the manufacturer's protocols (Jiancheng Bioengineering Ltd., Nanjing, China). Levels of activated Caspase 3, which reflected apoptosis activity, were detected by using commercial kits (Beyotime, China).
3
Wound-Healing Assay for Lung Cancer Cells
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The migratory capability of lung cancer cells was examined using wound-healing assay. Briefly, cells were seeded in 6-well plates and grown to full confluency, followed by overnight incubation in starvation medium. Cell monolayer was scratched with a 10 μL sterile pipette tip, washed with PBS, and then cultured in serum-free medium for 24h. Wound gaps were observed under Olympus BX41 microscope and photographed using a Qcolor5 digital camera.
4
In Vitro Plaque and Vascular Binding of PiB
5
Immunostaining Protocol for Cardiac Cell Lines and Tissue
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H9c2 cells were fixed with 10% formalin for 15 min at room temperature (RT). After this, the cells were permeabilized with 0.2% Triton X-100 in PBS for 10 min. Non-specific binding was blocked via incubation with 10% FCS in PBS for 30 min at 37 °C in a humidified chamber. H9c2 cells were immunostained with fibroblast-specific protein-1 (FSP-1) primary antibodies (1:200) followed by incubation with secondary antibodies conjugated with fluorescein isothiocyanate (FITC) or Cy3 (1:500). The area percentage of fluorescence intensity was quantified at six random microscopic fields using Image J analysis software (Image J, NIH, Bethesda, MD, USA).
Left ventricular samples were embedded and frozen in OCT compound (4583; Sakura Finetek, Torrance, CA, USA), and 10 µm frozen sections were prepared. Sections were immunostained with Troponin and FSP-1 primary antibodies (1:200) followed by incubation with secondary antibodies conjugated with fluorescein isothiocyanate (FITC) or Cy3 (1:500). Photomicrographs were obtained with an Olympus BX51 microscope, a Q-Color5 digital camera and a Q-Capture Suite acquisition software (Olympus, Tokyo, Japan).
Left ventricular samples were embedded and frozen in OCT compound (4583; Sakura Finetek, Torrance, CA, USA), and 10 µm frozen sections were prepared. Sections were immunostained with Troponin and FSP-1 primary antibodies (1:200) followed by incubation with secondary antibodies conjugated with fluorescein isothiocyanate (FITC) or Cy3 (1:500). Photomicrographs were obtained with an Olympus BX51 microscope, a Q-Color5 digital camera and a Q-Capture Suite acquisition software (Olympus, Tokyo, Japan).
6
Scratch Wound Healing Assay in Myoblasts
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The myoblasts were plated in four-chamber slides (Thermo Fisher Scientific) to 80% confluency in growth medium. A scratch line was drawn through the cell monolayer using a 200-μl pipette tip. Lifted myoblasts were washed away with growth medium and the medium was replaced with growth medium with paxilline or vehicle.19 (link) The myoblasts were cultured at 37 °C, 5% CO2, for 6 h and the scratch injury area was captured at room temperature using BX41 microscope (Olympus, Melville, NY, USA) equipped with Plan N × 10/0.25 objective lens and Qcapture software (Surrey, BC, Canada) linked to a Q-Color5 digital camera (Olympus). The area of scratch injury was measured using Adobe Photoshop CS3 (San Jose, CA, USA).
7
Perkinsus Cell Cycle Characterization
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Considering that different cell phases (trophozoites, schizonts and zoospores) of Perkinsus spp. develop on in vitro culture (Sunila et al.2001 (link); Casas et al.2002 (link); Dang et al.2015 (link)), light microscopy was used to correlate cell populations observed by this technique with those detected by flow cytometry.
The cell suspensions (4 replicates/treatment/time = 120 samples) were fixed in formaldehyde (final concentration of 2%) and used to identify and characterize (cell diameter and morphological description) each cell phase under a light microscope (Olympus BX45). Images of P. marinus cell cycle phases were taken with an Olympus Q-Color 5 digital camera.
The cell suspensions (4 replicates/treatment/time = 120 samples) were fixed in formaldehyde (final concentration of 2%) and used to identify and characterize (cell diameter and morphological description) each cell phase under a light microscope (Olympus BX45). Images of P. marinus cell cycle phases were taken with an Olympus Q-Color 5 digital camera.
8
In situ Localization of Terpenoids
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For each O3 exposure time (three, five or seven days), three leaves from the sixth oldest node were used. The medial region of fresh, fully expanded leaf blades was freehand sectioned. Five sections were used for in situ localization of terpenoids using Nadi reagent (naphthol+dimethyl-paraphenylenediamine) [50] . Sections were incubated in the dark for 60 min at room temperature in Nadi reagent, prepared immediately prior to staining. After incubation, the sections were rinsed for 2 min in a sodium phosphate buffer (0.1 M, pH 7.2). By oxidation this reagent forms indophenol blue that changes colour with variation in pH and makes it possible to distinguish between essential oils (blue) and resin acids (intense red), in which a purple color is observed when both compounds constitute the secretion [50] , [51] (link). Five other sections remained untreated – not submitted to any dye or reagents – for observation of the colour and structure of the cells in vivo. Observations and digital images were acquired with an Olympus BX53 compound microscope equipped with an Olympus Q-Color 5 digital camera and Image Pro Express 6.3 software.
9
Renal Cortex Histological and Immunohistochemical Analysis
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The renal cortex tissues were fixed with buffered 10% formalin solution (SF93–20; Fisher Scientific, Pittsburgh, PA, USA), embedded in frozen OCT compound (4583; Sakura Finetek, Torrance, CA, USA) and 10 μm frozen sections prepared. Some sections were stained with haematoxylin & eosin. Some were directly immunostained with Alexa 488‐labelled Isolectin B4 (IB4) for ECs, smooth muscle actin (SMA) and a NG2 antibody for pericytes (1:100; Abcam). Other sections were immunostained with Notch3, PHD2, FSP‐1 and TGF‐β1 primary antibodies (1:200) followed by incubation with second antibodies conjugated with fluorescein isothiocyanate (FITC) or Cy3 (1:500). Photomicrographs were obtained with an Olympus BX51 microscope, a Q‐Color5 digital camera and a Q‐Capture Suite acquisition software (Olympus, Tokyo, Japan). The area percentage of fluorescence intensity was quantified at six random microscopic fields using image analysis software (Image J, NIH, Bethesda, MD, USA). Masson trichrome staining was also performed to measure the degree of fibrosis (blue) in the renal cortex.
10
Melanoma Cell Migration Assay
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The migration ability of melanoma cells was determined in vitro using Boyden Chambers (Gaithersburg, MD) in which the two chambers were separated with Millipore membranes (8 μM pore size), as detailed previously [15 (link), 24 (link)]. The membranes were examined microscopically and cellular migration was determined by counting the number of stained cells on membranes in at least 3-4 randomly selected fields using an Olympus BX41 microscope with 10x magnification. Representative photomicrographs were obtained using a Qcolor5 digital camera fitted to an Olympus BX41 microscope. Cell migration experiments were repeated to verify the results.
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