Tissue punches (4 mm) were fixed and stored in 80% methanol at 4 °C. Tissues were cyroprotected in 10% (1 h), then 20% (6 h), and then 30% (overnight) sucrose. Samples were embedded in optimal cutting temperature medium (OCT, Thermo Fisher), sectioned at 14 µm and thaw mounted on glass slides. Samples were then stained with Safranin-O (Sigma-Aldrich). Sections were imaged using a Zeiss Axioskop 40 and images were taken with an AxioCam MR (Zeiss), and processed using AxioVision LE64 software (Zeiss).
Axiovision le64
Manufactured by Zeiss
Sourced in Germany
AxioVision LE64 is a microscope imaging software developed by Zeiss. It provides a platform for capturing, processing, and analyzing digital images from Zeiss microscopes.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1, by Zeiss (6 mentions)
Axioskop 2 plus, by Zeiss (6 mentions)
Axioskop2 mot plus wide field fluorescence microscope, by Zeiss (4 mentions)
Axiocam mr, by Zeiss (2 mentions)
Axioskop 40, by Zeiss (2 mentions)
Prolong gold antifade containing dapi, by Thermo Fisher Scientific (2 mentions)
Axiocam camera, by Zeiss (2 mentions)
Hbo illuminator, by Zeiss (2 mentions)
89 north photofluor lm 75 illumination system, by Chroma Technology (2 mentions)
Lsm 510 meta, by Zeiss (2 mentions)
Safranin o, by Merck Group (1 mentions)
M205 fa, by Leica (1 mentions)
Depex, by Serva Electrophoresis (1 mentions)
Axiocam icm1 camera, by Zeiss (1 mentions)
Bovine collagen type 1, by Advanced BioMatrix (1 mentions)
Axiovert 100 microscope, by Zeiss (1 mentions)
Illustrator cs6, by Adobe (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Dab detection ihc kit, by Abcam (1 mentions)
Safranin o fast green, by Merck Group (1 mentions)
22 protocols using axiovision le64
1
Tissue Cryosectioning and Safranin-O Staining
2
Transwell Migration Assay Protocol
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A 24-well plate was equipped with transwell migration assay inserts (8.0 µm pore size, Falcon, USA). The lower chambers were filled with 1ml culture medium containing 10% FCS whereas the upper chambers contained 0.8 mL culture medium without FCS. 200,000 cells were added to the upper chamber and cultured at 37 °C and 5 % CO2. After 48 h, inserts were removed, cells on the upper side were carefully removed with a cotton swap, and migrated cells on the lower side of the membrane were stained with Haema quick-stain kit. The membranes were cut out and mounted with mounting medium on glass slides (DePeX, Serva, Germany). Pictures of the whole membrane were taken using a stereo microscope (M205FA, Leica, Germany). Stained areas were analyzed by using the Carl Zeiss Axio Vision LE64 program.
3
Measuring MDA-MB-468 Cell Diameter
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MDA-MB-468 cell culture cells were trypsinized and resuspended in medium. The cell suspension was transferred to a glass slide via cytocentrifuge (190× g, 3 min). The resulting cytospins were left to dry overnight and stained with fluorescently labelled antibodies for pan-keratin and DAPI, as described above. Subsequently, 50 of the fluorescently labelled cells were photographed. The cell diameter was determined using the AxioVision LE64 microscope software (Zeiss) measurement tool and utilizing the borders of cytokeratin expression. An average was calculated from 50 separate cell measurements.
4
Collagen-Embedded Spheroid Invasion Assay
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Four similar size spheroids were obtained upon cell seeding in cell repellent 96-well round-bottomed plates. Spheroids were thereafter embedded into 2.7 mg/ml type I bovine collagen (Advanced BioMatrix, Carlsbad, CA). Once polymerized, the cell collagen matrix was overlayed with a 10% FCS-containing culture medium or NIH 3T3 cell supernatant. The invasion was documented upon image acquisition with an AxioCam ICm 1 camera coupled to an Axiovert 100 microscope and processed with the AxioVision LE64 program (Carl Zeiss, Oberkochen, Germany). The distance invaded by all of the 50 cells furthest from the spheroid center was measured using ImageJ software [67 ]. Median invaded distance was computed upon radius subtraction of a reference time point.
5
Fluorescent Microscopy Imaging Workflow
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Conventional fluorescent images and optical sections were captured with a Zeiss AxioObserver Z.1 inverse microscope with ApoTome function. Images were taken and processed with AxioVision LE64 (Carl Zeiss Microscopy GmbH, Jena, Germany), figures were assembled with Adobe Photoshop CS6 (Adobe Systems Incorporated, San José, CA, USA), and models were generated in Adobe Illustrator CS6 (Adobe Systems Incorporated). Charts were generated in Microsoft Excel 2016 (Microsoft Corporation, Redmond, WA, USA).
6
Microscopic Analysis of Loquat Fruit Cortex
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Three representative fruit samples from diverse loquat accessions, developmental phases, and treatments were fixed in formalin-acetic acid-alcohol (FAA) solution for at least 48 h. Cross-sections were obtained according to previously described methods15 (link). Then, the cortex cells of each section were observed under a light microscope with Axio Vision LE64 software (Carl Zeiss, Germany). To determine the effects of EjBZR1 on the cell size of Arabidopsis, petals of newly opened flowers were observed directly. The cell size in each sample was obtained using Image-Pro Plus 6 software (Media Cybernetics, America).
7
Radiographic Quantification of ECBB
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Antero-posterior (AP) and medio-lateral (ML) radiographs were taken and the ECBB around each collar was quantified in both the AP and the ML planes. ECBB resulted in a pedicle of bone that was measured from the shoulder of the collar using 3 different parameters; maximum thickness (mm), maximum length (mm) and surface area (mm 2 ). Measurements were calibrated on AxioVision LE64 software (v4.9.1.0, Carl Zeiss Microscopy GmbH, Oberkochen, Germany) using known diameters of the implant intramedullary stem prior to tissue analysis. ECBB seen on radiography did not confirm whether there was direct bone attachment with the implant. We analysed this using microscopy of histological thin sections.
8
IVD Tissue Characterization Protocol
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For intact IVDs, post-MRI analysis, a 2 mm wide sagittal tissue segment from the center of the IVD was fixed in periodate lysine paraformaldehyde (PLP) fixative overnight at 4°C. Samples were then washed in PBS and decalcified using Shandon TBD1 Decalcifier solution (ThermoFischer Scientific) over 72 hr at 4°C, changing solution each day. Tissue segments were washed in PBS and placed in 70% ethanol prior to paraffin embedding. Sections of 5 µm were cut and mounted on glass slides. All sections were heated on a hot plate at 55°C for 45 min and deparaffinized and rehydrated. Next, sections were stained with safranin-O/fast green (Sigma-Aldrich, Oakville, ON, Canada) and with antibodies against p16Ink4a and Ki-67 and counter stained using the DAB detection IHC Kit (ab64264, Abcam, Cambridge, MA) following the manufacturer’s instructions. All images were acquired using a Zeiss Axioskop 40 and an AxioCam MR (Zeiss) and processed using AxioVision LE64 software (Zeiss).
9
Morphological Characterization of Strain WLT07
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Strain WLT07 was morphologically studied on different media under different growth conditions. The strain was inoculated onto malt extract agar (MEA; Oxoid, Hampshire, UK), Czapek yeast extract agar (CYA; Difco), yeast extract sucrose agar (YES), oatmeal agar (OA; Difco), and creatine sucrose agar (CREA) at three points on 90-mm Petri dishes and incubated at 25 °C in the dark for 7 days. All media were prepared as described by Visagie et al. [26 (link)]. Additional CYA plates were incubated at 4 °C and 37 °C for 7 days in the dark. After incubation, diameters of colonies on each medium were measured. The density of sporulation, obverse and reverse colony colors, and the production of soluble pigments were noted. Fungal colonies were photographed with a Canon EOS 400 D camera (Tokyo, Japan). Morphological characterization was performed by observing the slides prepared from MEA using light microscopy (DE/Axio Imager.A1, Carl Zeiss, Göttingen, Germany). Lactic acid was used as a mounting fluid and a drop of ethanol was added to remove excess conidia. Specimen images were acquired using AxioVision LE64 software (Carl Zeiss, Oberkochen, Germany). Figure plates were prepared with Photoshop CS2 (Adobe, San Jose, CA, USA).
10
Immunofluorescence Microscopy of Parasites
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Parasites grown in HFF monolayers on glass coverslips were fixed in 4% (v/v) formaldehyde in PBS for 10 min, permeabilized by 0.25% (v/v) Triton X-100 in PBS for 20 min, and blocked in 3% BSA in PBS. Monolayers were incubated with different primary antibodies and visualized with secondary antibodies conjugated to Alexa Fluor. Coverslips were sealed onto slides using ProLong Gold Antifade containing DAPI (Thermo Fisher Scientific). Images were captured using a ×63 oil Plan-Apochromat lens (N.A. 1.4) on an Axioskop2 MOT Plus Wide Field Fluorescence Microscope (Carl Zeiss, Inc). Scale bars and linear adjustments were made to images using Axiovision LE64 software (Carl Zeiss, Inc).
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