The skin samples remaining after the completion of the (FITC)-dextran transport studies were subject to fluorescence microscopy analysis. To perform the analysis the skin samples were prepared by cutting them in half along their diameter and embedding them in O.C.T. compound. The embedded samples were sectioned in 20 μm slices using a cryostat microtome (Bright Instruments, Huntingdon, UK). The skin sections were mounted, stained with DAPI and covered with glass cover slips. Fluorescence photomicrographs were obtained with a Zeiss Axioscope microscope equipped with a Nikon Digital Camera (DXM1200; Nikon, Kingston upon Thames, UK) at a magnification of 10 ×. Images were acquired using two fluorescence channels to allow the visualization of the cellular structures stained by DAPI (blue colour emission ~ 460 nm) and the FITC-dextran fluorescence signal (green colour emission ~ 520 nm). Tissue samples without FITC-dextran were also tested as controls. Images were processed using Image J Software (National Institutes of Health, Maryland, USA).
Dxm1200 digital camera
Manufactured by Nikon
Sourced in Japan, United States, United Kingdom
The DXM1200 is a digital camera designed for scientific and industrial applications. It features a high-resolution CMOS sensor and a range of connectivity options for integration with various lab equipment and systems. The camera is capable of capturing detailed images and video for a variety of research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse e600 microscope, by Nikon (4 mentions)
Smz1000 dissection microscope, by Nikon (3 mentions)
Eclipse te200 microscope, by Nikon (2 mentions)
Act 1, by Nikon (2 mentions)
Biotinylated anti rabbit igg, by Vector Laboratories (2 mentions)
Abc vector elite, by Vector Laboratories (2 mentions)
Axioscope microscope, by Nikon (1 mentions)
Dm lb2 microscope, by Leica (1 mentions)
Bovine serum albumin (bsa), by Cell Signaling Technology (1 mentions)
Eclipse 80i fluorescent microscope, by Nikon (1 mentions)
E600 microscope, by Nikon (1 mentions)
Tcs sp8 x, by Leica (1 mentions)
Eclipse e600, by Nikon (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
Dm4000b microscope, by Leica (1 mentions)
Hyaluronidase, by Merck Group (1 mentions)
Hrp conjugated anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Axioplan 2 microscope, by Zeiss (1 mentions)
Anti pcna, by Agilent Technologies (1 mentions)
Anti βiii tubulin, by Promega (1 mentions)
51 protocols using dxm1200 digital camera
1
Fluorescence Microscopy Analysis of FITC-Dextran Skin Penetration
2
Morphological Changes in HeLa Cells
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To observe morphological changes, the HeLa cells were seeded in 35-mm dishes, cultured for 24 h and then treated with 40 μg/ml TAT-BID, 0.5 μM DOX or both of these agents together for 24 or 48 h, respectively. The cells were then washed twice with PBS and fixed with ice-cold methanol for 10 min at 4°C. Next, the cells were washed twice with PBS and stained with 0.25% May-Grünwald for 3 min at room temperature. After that 0.1 M phosphate buffer pH 7.0 was added (1:1) and left for 5 min. Next, the cells were stained with 0.76% Giemsa’s azur for 15 min at room temperature, washed at least three times and dried. Images (magnification, x100) were captured using a Nikon eclipse TE200 microscope equipped with a Nikon Digital Camera DXM 1200 (Nikon Instruments).
3
Microscopy analysis of lung tissue
4
Anchorage-independent Growth Assay
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Cells were resuspended in the agarose medium consisting RPMI-1640 medium, 10% v/v FBS, 0.3% w/v agarose, and delphinidin, and plated in a 6-well plate pre-coated with agarose (RPMI medium containing 10% v/v FBS and 0.6% w/v agarose). After incubated in a humidified atmosphere at 37 °C containing 5% CO2 for 7 days, the cells were fixed, stained with crystal violet, and then photographed under a microscope (Nikon Eclipse TE2000-U equipped with a Nikon Digital Camera DXM1200; Nikon, Japan). Colonies greater than 0.1 mm in diameter were counted.
5
Immunofluorescence Staining of MDA-MB-231 Cells
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After 48 h of treatment with Thiostrepton, MDA-MB-231 cells were washed with PBS and fixed in 4% paraformaldehyde for 30 min at room temperature. PBS with Tween-20 and 5% bovine serum albumin (Cell Signaling Technology, Inc., Danvers, MA, USA) was used to block the washed cells for 30 min at 37°C. The cells were incubated with primary antibodies at 4°C overnight and were subsequently stained with a fluorescent secondary antibody (Alexa-Fluor™ 594 donkey anti-rabbit IgG; 1:1,000; cat. no. A-21207; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 30 min in the dark. DAPI (cat. no. C1006; Beyotime Institute of Biotechnology, Haimen, China) was used to stain the nucleus for 15 min at room temperature. Immunopositive cells were observed under a fluorescence microscope. The sections were observed under a Nikon ECLIPSE 80i fluorescent microscope (Nikon Corporation; magnification, ×40) and the images were captured using a Nikon digital camera DXM1200 (Nikon Corporation).
6
Histopathological Evaluation of Knee Osteoarthritis
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Knee joints were removed and fixed in 4% paraformaldehyde in phosphate-buffered saline solution. Samples were decalcified in 10% EDTA, dehydrated in graded ethanol, cleared in toluene and embedded in paraffin. Serial sections were cut (7 μm) and mounted on SuperFrost glass slides (Menzel-Gläser, Braunschweig, Germany). Sections were deparaffinised in xylol, rehydrated in graded ethanol and stained with haematoxylin and eosin (general morphology) or safranin O (proteoglycan). The sections were mounted on a DPX medium (Panreac, Barcelona, Spain) and examined under a light microscope (Nikon Eclipse E800, Izasa S.A., Valencia, Spain) with 10x objective lens and a Nikon Digital Camera DXM1200 using Nikon ACT-1 software. Histopathological grading of cartilage degradation was performed on haematoxylin-eosin and safranin O stained sections by two independent, blinded observers in accordance with the Osteoarthritis Research Society International (OARSI) histopathology grading and staging system [22 (link)]. Synovitis was scored according to Krenn et al. [23 (link)].
7
Histological Analysis of Tissue Samples
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For histological analysis, samples were fixed in 10% neutral buffered formalin and then embedded into paraffin. Sections were cut at 5 μm, deparaffinized and stained with Heamatoxylin and Eosin (H&E), Safrannin-O (Saf-O). All histological results were analyzed with a digital image analysis system (Nikon E600 Microscope with a Nikon Digital Camera DXM 1200, Nikon Corporation, Japan).
8
Quantitative Lung Morphometry Analysis
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Lung morphometric analysis was performed as previously detailed44 (link). Briefly, mice were sacrificed and the left lung was inflated with 0.8% low melting agarose in 1.5% paraformaldehyde at 10–12 cm H2O pressure; this process was monitored continuously until agarose was solidified. Animal was placed at 4 °C for 30 min, the left lung was excised and kept in 1.5% paraformaldehyde for 24 h at 4 °C, washed with water and preserved in 70% ethanol. Lung was dissected into three pieces (2–3 mm thick) and embedded in paraffin, sectioned (5 μm) and stained with haematoxylin and eosin (H&E). Before morphometry, slides were coded and the whole lung images were acquired with Nikon Digital Camera DXM1200 (Nikon, Tokyo, Japan) or with an Aperio AT2® scannner (Leica) at ×10. The mean chord length (MCL) of digital JPEG images of the lung were evaluated using computer-assisted STEPanizer1 software45 (link) (www.stepanizer.com ) with sampling grid lines 17 μm apart, ensuring one to two chords per alveolus. Lung sections with arteries, veins, or bronchioles were excluded from the analysis.
9
Immunohistochemical Analysis of Liver Fibrosis
10
Microstructural Analysis of Cooked Samples
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Microstructural analysis was done before and after 240 min of digestion for all cooking methods. Samples were fixed in 10% Neutral Buffered Formalin for four days. After this time, samples were placed in 30% ethanol and processed on a Tissue Tek II Vacuum Infiltration Processor (Sakura Finetek, Alphen aan den Rijn, Netherlands) followed by embedding with paraffin using the ThermoFisher HistoCenter III embedding station (ThermoFisher Scientific, Waltham, MA, U.S.A.). Samples were cut in half and embedded with the cut surface down. Once samples were cooled, the excess of paraffin was removed from the edges. Samples were placed on a Reichert Jung 2030 rotary microtome (Reichert Technologies, Depew, NY, U.S.A.) and were finely sectioned at 5e6 mm.
Sections were air-dried overnight and then were placed in a 56 C slide incubator to ensure adherence to the slides for 2e24 h. Sections were stained with both Toluidine Blue (Mandalari et al., 2008) , and Periodic Acid Schiff (Tumuhimbise et al., 2009) . Samples were examined using a light microscope with 10 and 20Â objectives, and images were taken with a Nikon Digital Camera DXM1200 (Nikon Instruments Inc., Melville, NY, U.S.A.).
Sections were air-dried overnight and then were placed in a 56 C slide incubator to ensure adherence to the slides for 2e24 h. Sections were stained with both Toluidine Blue (Mandalari et al., 2008) , and Periodic Acid Schiff (Tumuhimbise et al., 2009) . Samples were examined using a light microscope with 10 and 20Â objectives, and images were taken with a Nikon Digital Camera DXM1200 (Nikon Instruments Inc., Melville, NY, U.S.A.).
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