Axiophot 2 microscope

Manufactured by Zeiss

Sourced in Germany, United States

The Axiophot 2 microscope is a high-performance optical microscope designed for a variety of applications. It features a motorized focus drive, a large mechanical stage, and a modular design that allows for the integration of various observation techniques, such as brightfield, darkfield, and fluorescence microscopy.

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Lab products found in correlation

Axiocam hrc camera, by Zeiss (6 mentions) Axiocam mrc, by Zeiss (6 mentions) Zen 2, by Zeiss (5 mentions) Neurolucida morphometry software, by MBF Biosciences (4 mentions) Alexa 555, by Thermo Fisher Scientific (4 mentions) Euparal, by Thermo Fisher Scientific (4 mentions) Vectashield with dapi, by Vector Laboratories (4 mentions) Alexa 488, by Thermo Fisher Scientific (4 mentions) Mouse anti v5, by Thermo Fisher Scientific (4 mentions) Neuroexplorer software, by MBF Biosciences (3 mentions) Fluoromount g mounting medium, by Electron Microscopy Sciences (3 mentions) Vectashield, by Vector Laboratories (3 mentions) Axiovision software, by Zeiss (2 mentions) Alexa 647, by Thermo Fisher Scientific (2 mentions) Rabbit anti gfp, by Abcam (2 mentions) Rabbit anti psmad3, by Abcam (2 mentions) Rabbit anti v5, by Abcam (2 mentions) Sp5 confocal microscope, by Leica camera (2 mentions) Lsm710, by Leica camera (2 mentions) Rat anti ollas, by Abnova (2 mentions)

67 protocols using axiophot 2 microscope

Microscopic Analysis of Retinal Cell Density

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Tissue was analyzed with a Zeiss Axiophot 2 microscope and a Zeiss Axioplan 2 equipped with epifluorescence. Micrographs were taken with a CCD camera and the Axiovision LE software (Carl Zeiss Vision, Germany). Some micrographs were taken with a laser scanning microscope Olympus FluoView 1000, using the FV 1.7 software (Olympus). The images were adjusted for brightness and contrast using Adobe Photoshop CS5.
To determine the density of X-gal-stained cells in the ganglion cell layer (GCL) and inner nuclear layer (INL) of flat-mounted retinae, counts were made in 250μm x 250μm sample fields with a 40x oil immersion objective, focused either on the GCL or the INL. Densities were not corrected for shrinkage, which was negligible in the tissues mounted with the aqueous medium.

Arbogast P., Flamant F., Godement P., Glösmann M, & Peichl L. (2016). Thyroid Hormone Signaling in the Mouse Retina. PLoS ONE, 11(12), e0168003.

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GUS Histochemical Staining for Time-Course and Developmental Analysis

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GUS staining was performed as previously described [16 (link)] with modifications [9 (link),12 (link)]. In detail, for time course analysis, after 50 μM MeJA treatment, samples were gently soaked in ice cold 90% (v/v) acetone for 30 min for prefixation, rinsed three times with sodium phosphate buffer (50 mM, pH 7.0) and then immersed in staining solution [1 mM 5-bromo-4-chloro-3-indolyl-β-d-glucuronide, 2.5 mM potassium ferrocyanide, 2.5 mM potassium ferricyanide, 0.1% (v/v) Triton X-100, 10 mM EDTA in sodium phosphate buffer (50 mM, pH 7.0)]. Histochemical GUS staining was allowed to proceed until differences in the intensity between treated and untreated plants were detected under the microscope (2 h). For developmental tissue-specific gene expression, the reaction proceeded overnight at 37 °C in dark. Chlorophyll was extracted by washing in sequence with ethanol/acetic acid ratio 1:3 (v/v) for 30 min, ethanol/acetic acid ratio 1:1 (v/v) for 30 min and with 70% ethanol for another 30 min. Samples were stored in 70% ethanol at 4 °C, prior to being observed under LM. Images were acquired by a Leica DFC450C digital camera applied to a Zeiss Axiophot 2 microscope.

Fraudentali I., Pedalino C., Tavladoraki P., Angelini R, & Cona A. (2021). A New Player in Jasmonate-Mediated Stomatal Closure: The Arabidopsis thaliana Copper Amine Oxidase β. Cells, 10(12), 3399.

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Quantifying Macrophages and T Cells in Nerve Tissue

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Ten-micrometer cryosections of the common peroneal/tibial nerve and the sural nerve were prepared using a cryostat (Leica, Blenheim, Germany). Immunohistochemical staining with antibodies against CD11b (rat, 1:250, Serotec, MCA711, Puchheim, Germany) for the detection of monocytes/macrophages (further referred to in the text as “macrophages”) and CD 3 (rat, 1:100, Serotec, MCA1477, Puchheim, Germany) for the detection of T cells were performed following standard methods using 0.02% diaminobenzidine (DAB) as chromogen and hemalaun as a counter staining. Anti-rat IgG were used as secondary antibodies. On negative control sections the primary antibody was omitted. Images were acquired using an Axiophot 2 microscope (Zeiss, Oberkochen, Germany) equipped with a CCD camera (Visitron Systems, Tuchheim, Germany). Immunopositive profiles were quantified manually in at least three sections for each mouse and related to the area of the sections. Data were analyzed using SPOT software (software version 5.2, Spot Software BV, Amsterdam, Netherlands) and ImageJ free software version 1.51f (National Institute of Health, Staten Island, NY, USA).

Karl F., Grießhammer A., Üçeyler N, & Sommer C. (2017). Differential Impact of miR-21 on Pain and Associated Affective and Cognitive Behavior after Spared Nerve Injury in B7-H1 ko Mouse. Frontiers in Molecular Neuroscience, 10, 219.

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Chromosome Preparation and Fiber-FISH Analysis

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Chromosome preparations, DNA fibers obtention, single and double-color FISH, and Fiber-FISH experiments were conducted as described in Kuhn et al. (2008) (link). The probes labeled with digoxigenin-11-dUTP were detected with antidigoxigenin FITC (Roche) and probes labeled with biotin-14-dATP were detected with NeutrAvidin-rhodamine (Roche). Chromosomes were stained with DAPI (4′, 6-diamidino-2-phenylindole, dihydrochloride salt). The preparations were analyzed under an epifluorescence Zeiss Axiophot 2 microscope equipped with a CCD camera and the images were obtained using the AxioVision software (Zeiss). To determine the size of the DNA fibers, hybridization signals were measured according to the protocol described by Schwarzacher and Heslop-Harrison (2000) .

de Lima L.G., Svartman M, & Kuhn G.C. (2017). Dissecting the Satellite DNA Landscape in Three Cactophilic Drosophila Sequenced Genomes. G3: Genes|Genomes|Genetics, 7(8), 2831-2843.

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Reconstruction of Hippocampal Pyramidal Neurons

Cited 3 times

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To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma42 (link)43 (link)44 (link). Neurons meeting these criteria were reconstructed in three-dimensions (3D) with a 40×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience). Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30 μm increments were analyzed in order to assess morphological cellular diversity and potential differences among animals45 (link).

Lazarczyk M.J., Kemmler J.E., Eyford B.A., Short J.A., Varghese M., Sowa A., Dickstein D.R., Yuk F.J., Puri R., Biron K.E., Leist M., Jefferies W.A, & Dickstein D.L. (2016). Major Histocompatibility Complex class I proteins are critical for maintaining neuronal structural complexity in the aging brain. Scientific Reports, 6, 26199.

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Dendritic Morphology Analysis of CA1 Pyramidal Neurons

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To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma
[75 (link), 76 (link), 78 (link)]. Neurons meeting these criteria were reconstructed in 3-dimensions (3D) with a 40×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience). Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30-μm increments
[79 (link)] were analyzed in order to assess morphological cellular diversity and potential differences between the animal groups.

Price K.A., Varghese M., Sowa A., Yuk F., Brautigam H., Ehrlich M.E, & Dickstein D.L. (2014). Altered synaptic structure in the hippocampus in a mouse model of Alzheimer’s disease with soluble amyloid-β oligomers and no plaque pathology. Molecular Neurodegeneration, 9, 41.

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Histological Examination of Abdominal Organs

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We also carried out a histological examination of the samples of abdominal organs (i.e., spleen, liver, small intestine, kidney) and of the related serous membranes (peritoneum and pleura). Additionally, we analyzed the large intestine and abdominal wall for the presence of the same alteration by collagenase lavage.
After fixing the different samples in 4% formalin at room temperature, these were numerically coded and transferred to the Department of Human Histology of Rey Juan Carlos University (Madrid, Spain) for analysis. The fixed tissues were embedded in paraffin and cut into 5-micron-thick slices. Sections were stained with hematoxylin–eosin. All were studied under a Zeiss Axiophot 2 microscope and photographed with an AxiocamHRc camera.

Garcia-Arranz M., Villarejo-Campos P., Barambio J., Garcia Gomez-Heras S., Vega-Clemente L., Guadalajara H, & García-Olmo D. (2022). Toxicity study in a pig model of intraperitoneal collagenase as an “enzymatic scalpel” directed to break stroma in order to generate a new perspective for peritoneal carcinomatosis approach: an experimental research. World Journal of Surgical Oncology, 20, 53.

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Quantitative Analysis of Hippocampal CA1 Neuron Morphology

Cited 3 times

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In order for a loaded neuron to be included in the analysis, it had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma (Radley et al., 2006 (link); Radley et al., 2008 (link); Dickstein et al., 2010 (link); Midthune et al., 2012 (link); Tyan et al., 2012 (link)). Neurons meeting these criteria were reconstructed in 3-dimension (3D) with a 40x/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience, Williston, VT). Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30-μm increments (Sholl, 1953 (link)) were analyzed in order to assess morphological cellular diversity and potential differences among animal groups.

Steele J.W., Brautigam H., Short J.A., Sowa A., Shi M., Yadav A., Weaver C.M., Westaway D., Fraser P.E., St George-Hyslop P.H., Gandy S., Hof P.R, & Dickstein D.L. (2014). Early Fear Memory Defects Are Associated with Altered Synaptic Plasticity and Molecular Architecture in the TgCRND8 Alzheimer's Disease Mouse Model. The Journal of comparative neurology, 522(10), 2319-2335.

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Immunostaining and Imaging of Drosophila Imaginal Discs

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The following primary antibodies were used: guinea-pig anti-Senseless (1:1000, gift from H. Bellen), mouse anti-Patched (1:50, Hybridoma bank), rabbit anti-V5 (1:500, Abcam), mouse anti-V5 (1:500, Invitrogen), rabbit anti-p-Smad3 (1:500, Epitomics), mouse anti-Dlp (1:50, Hybridoma bank), rabbit anti-GFP (1:500, Abcam), mouse anti-Wingless (1:200, Hybridoma bank). Secondary antibodies used were Alexa 488, Alexa 555 and Alexa647 (1:500, Molecular Probes). Total and extracellular immunostaining of imaginal discs was performed as previously described ⁴⁸. Imaginal discs were mounted in Vectashield with DAPI (Vector Laboratories) and imaged using a Leica SP5 confocal microscope. Confocal images were processed with ImageJ (N.I.H) and Photoshop CS5.1 (Adobe). All confocal images show a single confocal section. Adult wings were mounted in Euparal (Fisher Scientific) and imaged with a Zeiss Axiophot2 microscope with an Axiocam HRC camera. Adult wing size and L3-L4 intervein distance was measured with ImageJ.

Kakugawa S., Langton P.F., Zebisch M., Howell S., Chang T.H., Liu Y., Feizi T., Bineva G., O’Reilly N., Snijders A.P., Jones E.Y, & Vincent J.P. (2015). Notum deacylates Wnts to suppress signalling activity. Nature, 519(7542), 187-192.

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Immunohistochemical Analysis of Embryonic Smooth Muscle

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Embryos were harvested at 12.5 DPC and fixed in 20 volumes zinc-buffered formalin for 2h at room temperature, followed by overnight storage in 70% ethanol at 4°C, and processing through graded alcohols to paraffin. 5µm transverse sections were collected to positively charged glass slides and cured overnight at 42°C. Following de-waxing and rehydration, slides were re-fixed in Methyl Carnoy’s fixative, washed and incubated in 3% H2O2 to block endogenous peroxidases. Sections were washed in PBS, blocked for 1h in 1% BSA+ 10% normal goat serum at room temperature, followed by incubation in monoclonal anti-smooth muscle alpha actin antibody (DAKO), in the presence of 1% BSA + 2% normal goat serum, overnight at 4°C. Slides were next washed in PBS and incubated in a 1:200 dilution of HRP-conjugated goat ant-Mouse IgG (Jacksonimmuno) for 1h at room temperature, followed by signal color development via DAB with hydrogen peroxide (Vector Labs). Sections were counter-stained in Mayer hematoxylin and blued in Scott’s solution before dehydration and mounting with Cytoseal 60. Images were taken at 200x magnification on a Zeiss Axiophot2 microscope, equipped with a Zeiss Axiocam digital camera.

Donato A.J., Lesniewski L.A., Stuart D., Walker A.E., Henson G., Sorensen L., Li D, & Kohan D.E. (2014). Smooth Muscle Specific Disruption of the Endothelin-A Receptor in Mice Reduces Arterial Pressure, Vascular Reactivity and Affects Vascular Development. Life sciences, 118(2), 238-243.

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