Freshly lysed out RH‐GFP parasites (Kim et al, 2001 (link)) were filtered and diluted by ¼ in D10+ media. 2 ml of diluted parasite media was added to each coverslip and parasites were allowed to invade into the cells for 30 min at 37°C. After this, the cell monolayer was washed with PBS to remove uninvaded parasites. D10+ media alone or D10+ with DMSO, 40 µM combretastastin A4, or 40 µM parabulin was added to the dishes. Actively growing parasites in fibroblasts were imaged every 12 h over a 48‐h period using a Zeiss Axiovert 200 M and Axiovision camera to collect 10 random fields of view for each condition. Samples were stained with a 1:1,000 anti‐SAG1 mouse monoclonal antibody (Thermo Fisher) and detected with a 1:1,000 dilution of a goat anti‐mouse Alexa 594 secondary antibody (Thermo Fisher). Parasite replication states were manually quantified and tallied from 10 random microscope fields to compare the proportion of singlets, doublets, quadruplets, and 8+ rosettes. Data were displayed as the fraction of total vacuoles/field that contains singlet, doublet, quadruplet, or 8+ numbers. Three biological replicates with individual technical triplicates were quantified.
Axiovision camera
Manufactured by Zeiss
Sourced in Germany
The Axiovision camera is a high-performance digital camera designed for microscopy applications. It captures detailed, high-resolution images with accurate color representation. The camera's sensor and advanced imaging technology allow for efficient data acquisition and precise documentation of microscopic samples.
Automatically generated - may contain errors
Lab products found in correlation
Axiovert 200m, by Zeiss (4 mentions)
Axiovert microscope, by Zeiss (2 mentions)
35 mm coverslip dishes, by MatTek (1 mentions)
Axio imager a1, by Zeiss (1 mentions)
Sp 500 uz, by Olympus (1 mentions)
Picropodophyllin ppp, by Bio-Techne (1 mentions)
Recombinant igf 1, by R&D Systems (1 mentions)
Carl zoom inverted florescence microscope, by Zeiss (1 mentions)
Axiovision 4, by Zeiss (1 mentions)
Axio imager m1 microscope, by Zeiss (1 mentions)
Boyden chamber, by BD (1 mentions)
Alexa flour 488, by Thermo Fisher Scientific (1 mentions)
Stereo discovery v8, by Zeiss (1 mentions)
Quanta 250 feg, by Thermo Fisher Scientific (1 mentions)
10 protocols using axiovision camera
1
Imaging Parasite Replication Dynamics
2
Quantifying T. gondii Replication in HFF Cells
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HFF cells were grown to confluency in 35 mm coverslip dishes (MatTek, Ashland MA, USA). RH-GFP T. gondii [29 (link)] were harvested by filtering freshly lysed parasites. Parasites were diluted 4-fold in D10+ media. After 30 min of invasion at 37 °C, the cell monolayer was washed with PBS to remove extracellular parasites. D10+ media alone, D10+ with DMSO, or drug in DMSO was added to the dishes. Actively growing parasites in fibroblasts were imaged every 12 h over a 48-hour period using a Zeiss Axiovert 200 M and an Axiovision camera to collect 10 random fields of view for each condition. Parasite replication states were manually quantified and tallied to enumerate singlets, doublets, quadruplets, and larger rosettes. Data are displayed as the fraction of total vacuoles/field that contain singlet, doublet, quadruplet, or larger (8+) numbers at the 36-hour time point.
3
Optical Characterization of HFIB Spheroids
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To observe any possible variation in HFIB spheroids transparency and size after each optical clearing protocol is performed, optical microscopy images of the spheroids were acquired using a CX41 inverted optical microscope (Olympus, Hamburg, Germany) equipped with an Olympus SP-500 UZ digital camera and an Axio Imager A1 inverted microscope (Carl Zeiss, SMT, Inc., Oberkochen, Germany) equipped with an AxioVision camera. The transparency was checked by placing the spheroids on a crosshatched background, as previously described elsewhere [18 (link),19 (link),21 (link),32 (link)]. The influence of the optical clearing methods on the size of the spheroids was determined by measuring their diameter before and after the clearing procedure. For this analysis, the optical microscopy images were analyzed by using an image processing program designed for scientific purposes—ImageJ (National Institutes of Health, Bethesda, MD, USA) [36 ], as previously described in detail [35 (link)].
4
Isolation and Culture of Rat DRG Explants
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Embryonic rat dorsal root ganglia (DRGs) were isolated as previously described by Greaves et al. (34 (link)). Whole-ganglion explants were plated onto poly-D -lysine- and Matrigel-coated wells (BD Biosciences) in 48-well plates and incubated in DRG medium (high-glucose DMEM supplemented with 0.01% penicillin-streptomycin, 10% fetal calf serum). Positive controls were supplemented with nerve growth factor (NGF) (Bio-Rad, Hercules, CA, USA) plus recombinant IGF-1 (R&D Systems, Minneapolis, MN, USA) (2–200 ng/ml) ± 125–500 nM Picropodophyllin (PPP; Tocris Bioscience, Bristol, United Kingdom). Some DRGs were incubated in PF or macrophage-conditioned medium diluted 1:1 in DRG medium ± 500 nM PPP. Images of explants were captured using an Axiovert microscope (Carl Zeiss, Oberkochen, Germany), an Axiovision camera, and software.
5
Oil Red O Staining for Lipid Quantification
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Cells were seeded in 6-well plate in triplicate and treated with 5 mmol/L aspirin with or without Compound C pre-treatment. After 24 h, the culture media was removed and cells were washed three times with PBS. Next, the cells were fixed with 4% paraformaldehyde at room temperature for 30 min, and rinsed with PBS once for 1 min. Thereafter, the cells were washed with 60% isopropanol for 15 s. Freshly diluted Oil Red O working solution (Hat Biotechnology, Xian, China) was then added, and the cells were incubated for 60 min at room temperature. Then, cells were rinsed again with 60% isopropanol for 15 s and washed three times with PBS for 5 min each. All images were captured using an Axiovision camera with a Carl Zeiss zoom inverted florescence microscope at a magnification of 60 ×. For the quantitation of lipid loading, the cells were seeded in 96-well plates and treated as described above. After removing the staining solution, the dye retained in the cells was eluted with isopropanol and the optical density at 540 nm (OD540) was determined with a spectrophotometer (Bio-Rad Laboratories).
6
Microscopic Image Acquisition and Editing
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Single or mosaic digital photographs of each section were made by using a Zeiss AxioImager M1 microscope mounted with an AxioVision camera and the AxioVision 4.6 software (Zeiss, Göttingen, Germany) and saved as 8-bit TIFF files. The acquired image files were opened in photo-editing software (Adobe Photoshop CS5), composed (cropped, rotated) and adjusted for brightness, contrast, or color balance.
7
Histological Analysis of Regenerated Tissue
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The samples were moved to plastic holders with 10% buffered formalin. The regenerated tissue and the adjacent native bone were divided into upper, middle, and lower segments. The samples were taken out from the 10% formalin, stacked into cassettes of appropriate size, and put in a rotor basket buffered in 10% formic acid for decalcification. Fluoroscopy was used to verify the end point of the decalcification procedure in order to prevent undue tissue injury. Sections of 5-µm were prepared from the decalcified tissues, which was immersed in paraffin wax to form blocks. Haematoxylin and eosin (H & E) and Masson trichrome stains were used to stain the sections. Slides were then studied under a light microscope (Olymp us, Japan) (Zeiss, Germany) supplemented with an Axio-Vision camera.
8
Boyden Chamber Cell Migration Assay
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Cells were seeded at a density of 1 × 105 cells/mL in the upper surface of a Boyden Chamber (size of the pores, 8 µm) (BD Biosciences, Franklin Lakes, NJ, USA) containing 0.7 mL of media (1% FBS +/− 100 ng/mL of soluble RANKL). The chamber was immersed in a 24-multiwell plate containing 0.7 mL of complete media (DMEM + 10% FBS for HOS cells or RPMI + 5% FBS for MOS-J cells). After 24 h of incubation at 37 °C, the cells on the upper filter that had not migrated through were removed by wiping with a cotton bud. The remaining cells were fixed in 1% (v/v) glutaraldehyde solution for 10 min at room temperature then washed twice with PBS (pH 7.2). Cells that had migrated through the filter were stained for 20 min using a 10% (v/v) purple crystal solution in water. After drying, the cells were visualized and counted in five microscopic fields using an AxioVision Camera (Zeiss, Marly-le-Roi, France). The number of migrated cells was measured using Image J software (National Institutes of Health, Bethesda, MD, USA) and was evaluated in three independent experiments (n = 6).
9
Quantifying GLUT4 and pSTAT3 in C2C12 Cells
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After overnight IL-15 experimental treatment, the C2C12 cells were fixed with ice cold methanol. Cells were blocked with 5% BSA in PBS-Tween followed by a series of washes. Cells were incubated overnight in a humidified chamber with primary antibodies (GLUT4 or pSTAT3) at 4°C. Alexaflour-488 (Molecular Probes) secondary antibodies were used and DAPI was used as a nuclear stain. Cells were imaged using a Zeiss inverted microscope and images were captured with an Axiovision camera. Quantification of GLUT4 on the plasma membrane and co-localization of STAT3 to the nuclei was performed using Image J and Fiji software (Schindelin et al., 2012 (link); Schneider et al., 2012 (link)).
10
Surface Analysis of Urine-Exposed Samples
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Observations of the samples’ surfaces were performed using a Zeiss SteREO Discovery.V8 stereoscopic microscope with an AxioVision camera (Zeiss, Oberkochen, Germany), and a scanning electron microscope (SEM) (Quanta 250 FEG, Thermo Fisher Scientific, Hillsboro, OR, USA). The samples were observed at the initial state and after exposure to artificial urine.
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