The samples were fixed in 10 % neutral buffered formalin, decalcified in Shandon TBD-2 Decalcifier (TBD; Thermo Scientific, Kalamazoo, MI, USA) and embedded in paraffin. Cartilage sections (5 μm thick) were stained with safranin O-fast green using an established protocol [50 (link)] to detect the glycosaminoglycan (GAG) positive matrix that is typical for the hyaline cartilage and normally stains red. Light microscopy images were taken at 4× magnification with Olympus BX63 microscope (Olympus, Japan) equipped with Olympus DP72 CCD camera using CellSens Dimension imaging software (Olympus, Japan). The images were analysed using Image J software. Previously described image processing protocol was adapted to quantify the amount of GAG distribution in each section [51 (link)]. Briefly, red colour intensity (RCI) was calculated in the entire cross-sectional slice area by obtaining red, green, and blue (RGB) image planes in a scale of 256 values (black = 0). The fraction of red (RF) was defined as the ratio of the R component to the sum of the R, G, and B components: RF = R/(R + G + B) and expressed as a percentage. Sections were additionally evaluated by 3 blinded observers using histological-histochemical grading system proposed by Mankin et al. [52 (link)]. Fresh articular cartilage received the score of 0, whereas higher score indicated more deteriorated cartilage.
Cellsens dimension imaging software
Manufactured by Olympus
Sourced in Japan, Germany, United States
CellSens Dimension is an imaging software designed for microscopy applications. It provides tools for image acquisition, processing, and analysis. The software supports a range of microscopy techniques and enables users to capture, manage, and analyze digital images.
Automatically generated - may contain errors
Lab products found in correlation
Dp72 ccd camera, by Olympus (2 mentions)
Dp72 digital camera, by Olympus (2 mentions)
Giemsa solution, by Carl Roth (2 mentions)
Roti histokitt mounting solution, by Carl Roth (2 mentions)
May gr nwald solution, by Carl Roth (2 mentions)
Bx63 fluorescence microscope, by Olympus (2 mentions)
Calcein am, by Thermo Fisher Scientific (2 mentions)
Ix73 inverted fluorescence microscope, by Olympus (2 mentions)
Bx43 microscope, by Olympus (2 mentions)
Bx53 microscope, by Olympus (2 mentions)
Shandon tbd 2 decalcifier, by Thermo Fisher Scientific (1 mentions)
Bx63 microscope, by Olympus (1 mentions)
Fv1000 inverted confocal ix81 microscope, by Olympus (1 mentions)
Orca flash 4.0 v3, by Olympus (1 mentions)
Fv10 asw 3, by Olympus (1 mentions)
Autoquant x3, by Media Cybernetics (1 mentions)
Colorview, by Olympus (1 mentions)
Sdf plapo 1xpf objective, by Olympus (1 mentions)
Dp72 camera, by Olympus (1 mentions)
Sytox green nucleic acid stain, by Thermo Fisher Scientific (1 mentions)
29 protocols using cellsens dimension imaging software
1
Quantifying Cartilage GAG Content
2
Olympus Confocal Microscopy Imaging
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Z-stack 3D images shown in Fig. 2a, b were acquired at 0.3-µm increments using an Olympus FV1000 Inverted Confocal IX81 Microscope and FV10-ASW 3.1 Software (Olympus). All other images were acquired using the Olympus IX83 fluorescence microscope system. Optical Z-sections were collected at 0.30- or 0.60-µm increments with the Hamamatsu Orca Flash 4.0 v3 and CellSens Dimension imaging software (Olympus). Pictures were deconvolved using AutoQuant X3 (Media Cybernetics).
3
Fluorescence Microscopy of Cryosections
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Cryosections and cells were examined using a light microscope (BX-53, Olympus LRI Instruments AB, Lund, Sweden) equipped with a mercury lamp and filters for fluorescence (U-MWG, U-MWB, U-MWU). Images were taken in 10x, 20x or 40x magnification using an Olympus Color View digital camera and captured using cellSens Dimension imaging software (Olympus).
4
Pig TMJ Zonal Collagen Analysis
5
Time-lapse Imaging of Early Cell Divisions
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One-cell embryos were microinjected as already described and allowed to develop at room temperature. Time-lapse imaging was performed with an Olympus stereomicroscope (Olympus SZX16 research fluorescence stereomicroscope equipped with Olympus DP72 camera, 11.5× zoom microscope body, and SDFPLAPO1XPF objective) at room temperature on the animal pole starting at 4 hpf. Images were acquired every 5 minutes. Discontinuous light was used to illuminate embryos, controlled with a digital adjustable cycle timer (CT-1 Short Cycle Timer, Innovative Grower Corp). The number of time intervals between cell divisions was counted to determine cell cycle lengths. Division timing was measured for at least 5 cells per embryo and for each cleavage. The diameter of surface-exposed cells was measured using cellSens Dimension imaging software (Olympus).
6
Quantifying Nuclei Morphometrics in Embryos
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Embryos previously stained by in situ hybridization were washed 3 × 1 hour at room temperature in TBST (TBS + 0.1% Tween 20), incubated overnight at 4 °C with Sytox Green nucleic acid stain (Life Technologies, S7020, 1:1000 dilution in TBST), and washed 3 × 1 hour in TBST at room temperature. Embryos were dehydrated in methanol, cleared in 2:1 benzyl benzoate:benzyl alcohol, and imaged in circular ~1.5 mm deep chambers covered with 25 × 25 mm glass coverslips. Images of nuclei were acquired using the same exposure time with an Olympus BX51 fluorescence microscope, Olympus UPLFLN 20x (N.A. 0.50, air) objective, and QIClick Digital CCD Camera (model QIClick-F-M-12, mono, 12-bit). DNA staining intensity and nuclear cross-sectional area were quantified from original thresholded images using cellSens Dimension imaging software (Olympus).
7
Nephrotoxic Effects of Viper Venoms
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The nephrotoxic effects of D. russelii and D. palaestinae venoms were assessed by injecting 10 μg of the venom into the caudal vein of 4 male CD-1 mice (18–22 g). Both kidneys were harvested immediately after the death of the animal and were washed with 1X PBS, fixed in 10% buffered formalin for 24 h, and dehydrated with ascending concentrations of ethyl alcohol (70 and 95% for 30 min; 100% for 2 h), and cleared in xylene (Thermofisher, Waltham, MA, USA). Tissues were embedded in paraffin (Thermofisher, Waltham, MA, USA) at 58 °C, following which 3 μm sections were prepared using a Leica microtome (RM2245, Wetzlar, Germany). The slides were then stained with hematoxylin (Leica, Wetzlar, Germany), eosin (Leica, Wetzlar, Germany) and Masson’s trichrome staining (MTS; Path Stains, Bengaluru, India). The slides obtained were visualised using an Olympus light microscope (Ix81, Olympus, Shinjuku, Japan) at a 40× magnification, and images were acquired and analysed using CellSens dimension imaging software (Olympus, Shinjuku, Japan). The histological structure of renal tubules and glomeruli of the treatment group (10 μg of venom) was assessed in comparison to the control that received 200 μL of normal saline [82 ].
8
Bacterial Motility Assessment by Microscopy
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Bacterial strains were grown to exponential phase and 10 μL of bacterial suspension adjusted at 106 bacteria mL−1 were prepared for dark-field contrast microscopy using a #1.5H square coverslip. Motility was assessed in liquid medium by video microscopy using a BX53 Olympus microscope (Olympus Corporation, Nishi-Shinjuku, Tokyo, Japan) equipped with a Hamamatsu 2.8 Orca flash camera (Hamamatsu Photonics, Hamamatsu City, Japan). For each strain, three experiments were performed on different days and using independent cultures. In total, 190 bacteria per strain were recorded over 60 s. Trajectory analysis and speed displacement were calculated using Olympus CellSens Dimension imaging software.
9
Immunohistochemical Staining of Retinal ADPase
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After 24 h of incubation in 4% PFA, the retinas were washed in tris-maleate buffer (pH 7.2) on ice prior to incubation in an ADPase incubation medium containing 3.0 nM of lead nitrate and 6.0 mM of magnesium chloride (Sigma Chemical Co., St. Louis, MO, USA) in tris-maleate buffer (pH 7.2). After incubation, the retinas were washed with tris-maleate buffer prior to the addition of diluted ammonium sulfide (Fisher Scientific, Silver Spring, MD, USA) for 1 min. The retinas were washed again in tris-maleate buffer and flatmounted on a microscope slide with PBS and glycerin. All of the images were captured using an Olympus BX53 microscope, DP72 digital camera, and CellSens Dimension imaging software (version 2.1) from Olympus America, Inc. (Center Valley, PA, USA), attached to a HP Z44 computer.
10
May-Grünwald Giemsa Staining Protocol
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A total of 4 × 104 cells were resuspended in 150 μL of phosphate-buffered saline (PBS) and centrifuged on glass slides using a Cytofuge® (Medite, Burgdorf, Germany) at 700× g for 10 min. Afterwards, May–Grünwald Giemsa staining was performed. In short, cells were stained in 0.25% (w/v) May–Grünwald solution (Roth, Karlsruhe, Germany) for 5 min, washed in distilled water and stained with 1:20 diluted GIEMSA solution (Roth) for 20 min. After a second wash step, the slides were dried overnight and embedded in Roti-Histokitt mounting solution (Roth, Karlsruhe, Germany). Pictures were taken using an Olympus IX71 with the CellSens Dimension imaging software (Olympus, Shinjuku, Japan).
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