Axiocam hr digital camera

Manufactured by Zeiss

Sourced in Germany

The Axiocam HR is a high-resolution digital camera developed by Zeiss. It is designed for use with microscopes to capture detailed images for various scientific and research applications.

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11 protocols using axiocam hr digital camera

Whole-Mount In Situ Hybridization Protocol

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Examination of gene expression by whole-mount in situ hybridization was performed essentially as previously described [33 (link)–37 (link)]. Prior to mRNA in situ hybridization analyses, embryos were fixed in 4% paraformaldehyde (PFA)/phosphate-buffered saline (PBS) overnight at 4°C or 4–5 hours at RT with gentle agitation on a rotating platform. Embryos were permeabilized in 10 μg/ml proteinase K for 10 seconds (10–12 hpf embryos), 30 seconds (14–17 hpf embryos), 3 minutes (24–32 hpf embryos), or 1 hour (3–4 days post fertilization embryos) at RT.
Following in situ hybridization, embryos were manually deyolked, and cleared in 30%, 50%, and 70% glycerol/PBS. Mounted in situ hybridized embryos and live Tg(gata1:DsRed)^sd2Tg embryos were photographed using a Zeiss AxioImager Z1 compound microscope with an Axiocam HR digital camera. Mounted Tg(kdrl:GFP)^la116Tg embryos were photographed using a Zeiss LSM510 confocal microscope. Whole embryos were photographed using an Olympus stereoscope with a QImaging micropublisher camera. Images were assembled in ImageJ or Zen (Zeiss), and figures were assembled in Photoshop (Adobe).

Pillay L.M., Mackowetzky K.J., Widen S.A, & Waskiewicz A.J. (2016). Somite-Derived Retinoic Acid Regulates Zebrafish Hematopoietic Stem Cell Formation. PLoS ONE, 11(11), e0166040.

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Fluorescence Microscopy Imaging Pipeline

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The mounted sections were scanned on a Zeiss Axio Imager.Z1 upright fluorescence microscope with a motorized stage and an Axiocam HR Digital Camera (Zeiss). A tiled mosaic image of all of the sections was obtained with a 10× objective using the MosaiX module of the Axiovision software (Zeiss). The MosaicPlanner software, written by Forrest Collman in the laboratory of Stephen Smith at Stanford (now at the Allen Brain Institute), was used to identify and mark the x–y coordinates of processes of the two filled neurons. The MosaicPlanner software is available for download at https://code.google.com/p/smithlabsoftware/. For high resolution images, a 5 × 8 tiled mosaic was taken at each section using these x,y coordinates with a 63×/1.4 NA Plan Apochromat objective with the MosaiX module for Axiovision. The mosaic images generated were digitally stitched together using the Axiovision MosaiX plugin using the DAPI as the reference channel. The stitched images were exported as TIFF files then aligned with ImageJ (NIH) using a custom plugin written by Forrest Collman, using the DAPI channel as a reference (https://code.google.com/p/smithlabsoftware). The images from all seven of the ribbon arrays were ordered into one large stack and aligned with the TrakEM2 plugin for ImageJ to remove non-linear warping.

Valenzuela R.A., Micheva K.D., Kiraly M., Li D, & Madison D.V. (2016). Array tomography of physiologically-characterized CNS synapses. Journal of neuroscience methods, 268, 43-52.

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Quantifying Marine Microbial Cells Using Epifluorescence Microscopy

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Freshly sampled sediments (0.5 cm³) were placed into vials with 14 mL of a 2% glutaraldehyde solution in ultrafiltered seawater and stored at +4 °C. In the laboratory, the volume of the suspension was adjusted to 50 mL with ultrafiltered seawater. The sample was sonicated on a UZV-2/150-TN-RELTEC device (Russia) under the following conditions: sample processing time 4 min, amperage 0.44 A, frequency 15 kHz. After desorption and precipitation of “heavy” particles, 0.5 mL of the suspension was filtered on black polycarbonate filters (Millipore) with a pore diameter of 0.2 µm. Filters were stained with acridine orange solution [33 (link)]. The preparations were examined using an epifluorescence microscope (Carl Zeiss, Germany) equipped with an Axio CamHR digital camera at a magnification of × 1000. Cells were counted from a monitor screen in 20 fields of view.

Begmatov S., Savvichev A.S., Kadnikov V.V., Beletsky A.V., Rusanov I.I., Klyuvitkin A.A., Novichkova E.A., Mardanov A.V., Pimenov N.V, & Ravin N.V. (2021). Microbial Communities Involved in Methane, Sulfur, and Nitrogen Cycling in the Sediments of the Barents Sea. Microorganisms, 9(11), 2362.

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Neutrophil Migration in 3D Collagen Gels

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The analysis of migration in collagen gels was performed in μ-slide chemotaxis chambers (IBIDI, Planegg, Germany). A gel–cell mixture consisting of 3 × 10⁵ neutrophils in 1.5 mg/mL type I rat tail collagen (IBIDI) was applied to the middle channel of the 3D chamber and left at 37°C for 5 min for gelation. After application of 100 nM fMLP for 20 min at 37°C, time-lapse videos were recorded for 10 min using an Axiovert 200 M microscope (Zeiss, Jena, Germany) equipped with a Plan-Apochromat 10×/0.75NA objective, AxioCam HR digital camera, and a temperature-controlled environmental chamber. Migration tracks were analyzed offline with the Image J software. Single cell migration tracks and rose plots were generated using the IBIDI Chemotaxis software.

Gonçalves-de-Albuquerque C.F., Rohwedder I., Silva A.R., Ferreira A.S., Kurz A.R., Cougoule C., Klapproth S., Eggersmann T., Silva J.D., de Oliveira G.P., Capelozzi V.L., Schlesinger G.G., Costa E.R., Estrela Marins R.D., Mócsai A., Maridonneau-Parini I., Walzog B., Macedo Rocco P.R., Sperandio M, & de Castro-Faria-Neto H.C. (2018). The Yin and Yang of Tyrosine Kinase Inhibition During Experimental Polymicrobial Sepsis. Frontiers in Immunology, 9, 901.

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Microscopy Imaging and Editing Protocols

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Histological sections were examined under an Axioskop 2 MOT microscope (Carl Zeiss, Göttingen, Germany). Micrographs were captured with an Axiocam HR digital camera (Carl Zeiss) and the KS400 image analysis software (v3.0, Carl Zeiss). Following shading correction in KS400, images were cropped, resized, and adjusted for brightness (including slight gamma changes), colour balance, and sharpness in Corel Photo-Paint X4. All adjustments were applied to the whole image and no specific features within the photographs were modified, removed, or inserted.

Washausen S, & Knabe W. (2018). Lateral line placodes of aquatic vertebrates are evolutionarily conserved in mammals. Biology Open, 7(6), bio031815.

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Chemotaxis Assay of pTr Cells

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The pTr cells (50,000 cells per 100 µl serum and insulin-free DMEM) were seeded on 8-µm pore Transwell inserts (Corning Costar no. 3422; Corning, Inc., Corning, NY). Cells were then treated with recombinant porcine CSF2 protein (0, 0.1, 1, 20 and 100 ng/ml)(n = 3 wells per treatment). After 12 h, cells on the upper side of the inserts were removed with a cotton swab. For evaluation of cells that migrated onto the lower surface, inserts were fixed in 50% ethanol for 5 min. The transwell membranes were then removed, placed on a glass slide with the side containing cells facing up, overlaid with prolong antifade mounting reagent with 4′,6- diamidino-2-phenylindole, and overlaid with a coverslip (Invitrogen- Molecular Probes, Eugene, OR). Migrated cells were counted systematically in five non-overlapping locations, which covered approximately 70% of the insert membrane growth area, using a Zeiss Axioplan 2 fluorescence microscope with Axiocam HR digital camera and Axiovision 4.3 software (Carl Zeiss Microimaging, Thornwood, NY). The entire experiment was repeated at least three times.

Jeong W., Kim J., Bazer F.W, & Song G. (2014). Proliferation-Stimulating Effect of Colony Stimulating Factor 2 on Porcine Trophectoderm Cells Is Mediated by Activation of Phosphatidylinositol 3-Kinase and Extracellular Signal-Regulated Kinase 1/2 Mitogen-Activated Protein Kinase. PLoS ONE, 9(2), e88731.

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Intestinal Morphometry: Histological Analysis

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Duodenum, jejunum, and ileum segments were fixed in 4% paraformaldehyde (pH 7.2) solution for 24 h, dehydrated and embedded in paraffin. Each selected sample was sectioned into six μm thickness with a microtome. The sections of the intestine were stained with hematoxylin and eosin. Using an image processing system composed of CMOS (OLYMPUS, Japan) and its special software (Imaging Technology, USA), the computer obtains the villus height, crypt depth, and ratios. Villus height and crypt depth were measured at × 40 magnification using a microscope (BA400Digital, Mike Audi Industrial Group Co., Ltd. Xiamen, China). The images were evaluated by using an Axioplan 2 microscope (Carl Zeiss, Thornwood, NY) interfaced with an Axiocam HR digital camera.

Liu Y., Wang D., Zhao L., Zhang J., Huang S, & Ma Q. (2022). Effect of Methionine Deficiency on the Growth Performance, Serum Amino Acids Concentrations, Gut Microbiota and Subsequent Laying Performance of Layer Chicks. Frontiers in Veterinary Science, 9, 878107.

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Histological Analysis of Muscle Atrophy

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For haematoxylin and eosin stain, fresh muscle sections were placed in haematoxylin for an appropriate time, no longer than 15 min, and briefly washed in distilled water. Sections were subsequently bled in 70% ethanol/0.1% hydrochloric acid for 10 s, and washed in distilled water. The sections were then placed in eosin for 2 min, washed in distilled water, dehydrated and then mounted using Histomount. For muscle atrophy measurements, images were acquired using an AxioCam HR digital camera (Zeiss) and the area of muscle in atrophy and total muscle area were obtained using AxioVision software (Zeiss).
For succinic dehydrogenase staining, fresh muscle sections were incubated in 0.05 M phosphate buffer/0.05 M sodium succinate/0.05% nitro blue tetrazolium chloride for 30 min at 37°C, and subsequently rinsed in PBS. Sections were then fixed in 4% PFA for 5 min and rinsed in 15% ethanol before being mounted using Histomount. All quantification was conducted with the experimenter blind to the genotypes.

Liu K.X., Edwards B., Lee S., Finelli M.J., Davies B., Davies K.E, & Oliver P.L. (2015). Neuron-specific antioxidant OXR1 extends survival of a mouse model of amyotrophic lateral sclerosis. Brain, 138(5), 1167-1181.

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Sporozoite Membrane Integrity Assay

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Nine-thousand P. gallinaceum mature sporozoites were recovered from the salivary glands of Ae. aegypti and incubated in 50 μL of the PBS solution, with 40 μmol L⁻¹ digitonin (positive control), 60 μmol L⁻¹ peptides or negative controls, at 37 °C for 1 h. Cell membrane integrity was then monitored using a Carl Zeiss inverted fluorescence microscope (model Observer Axio Vision A.1) coupled to an image capture Zeiss AxioCam HR digital camera (1300 × 1030 pixels resolution and 8-bit quantization) after addition of 1 μL of the propidium iodide aqueous solution (200 μmol L⁻¹) in 5 μL of total solution volume. Images were obtained using a 40× objective lens and a green filter effect in red. The spectral range was set with the excitation at 538 nm within the visible spectrum in order to produce orange-red fluorescence centered at 619 nm, which was processed using the Axio 4.7 software.

Silva A.F., Torres M.D., Silva L.D., Alves F.L., Pinheiro A.A., Miranda A., Capurro M.L, & Oliveira VX J.r. (2015). New linear antiplasmodial peptides related to angiotensin II. Malaria Journal, 14, 433.

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Immunohistochemical Analysis of Ovarian Proteins

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Ovaries were fixed in 4% buffered paraformaldehyde (PFA) (6 g PFA, 325 mg NaOH, and 15 ml 10 × PBS in 100 ml diethylpyrocarbonate-treated double distilled H₂O, pH 7.2) for 1 h at 4 °C and processed using standard procedures (Kerr et al., 2006 (link)). Paraffin sections (5 μm) were used for immunohistochemical (IHC) localization of proteins using a Vectastain Elite ABC kit (Vector Labs, Burlingame, CA) as previously described (Lee et al., 2012a (link); Lee et al., 2012b (link)) and according to the manufacturer’s protocols. The tissue sections were incubated with primary antibodies for p-AKT, p-ERK, p53, p27, SOD-2, caspase-3, BAX and X-linked Inhibitor of Apoptosis Protein (XIAP) at specific concentrations (as indicated in Table 1). The sections were further incubated with the secondary antibody (biotinylated IgG) for 45 min at RT. Negative controls included serum or IgG from the appropriate species related to the primary antibody at the same dilution. Digital images were captured using a Zeiss Axioplan 2 research microscope with an Axiocam HR digital camera (Carl Zeiss). The intensity of staining for each protein was quantified using Image-pro Plus as described previously (Lee et al., 2012a (link); Lee et al., 2012b (link)) according to the manufacturer’s instructions (Media Cybernetics, Inc., Bethesda, MD).

Sivakumar K.K., Stanley J.A., Arosh J.A., Pepling M.E., Burghardt R.C, & Banu S.K. (2014). Prenatal exposure to chromium induces early reproductive senescence by increasing germ cell apoptosis and advancing germ cell cyst breakdown in the F1 offspring. Developmental biology, 388(1), 22-34.

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