Axiocam hrc camera

Manufactured by Zeiss

Sourced in Germany, United Kingdom, France, Japan, United States, Canada

The AxioCam HRc is a high-resolution digital camera designed for microscopy applications. It features a 12.6-megapixel CMOS sensor capable of capturing high-quality images with a resolution of 4096 x 3072 pixels. The camera is designed to provide accurate color reproduction and fast data transfer rates, making it suitable for a variety of microscopy techniques.

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318 protocols using axiocam hrc camera

Microscopy Techniques for Imaging Specimens

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Whole-mount images were taken using a Leica MZFLIII microscope (Leica Microsystems, Wetzlar, Germany) fitted with a QImaging MicroPublisher 5.0 RTV camera and QCapture Pro 6.0 software (QImaging, Surrey, BC, Canada); a Zeiss AxioSkop2 microscope fitted with a Zeiss AxioCam HRc camera and Zeiss AxioVision Rel. 4.8 software (Carl Zeiss, Oberkochen, Germany); an Olympus MVX10 microscope (Olympus Corporation, Tokyo, Japan) fitted with a Zeiss AxioCam HRc camera and Zeiss AxioVision Rel. 4.8 software; an Olympus 1 × 71 inverted microscope fitted with a Hamamatsu ORCA-R2 monochrome camera and HCImage software (Hamamatsu Photonics, Hamamatsu, Japan); a Zeiss LSM 710 confocal microscope with Zeiss ZEN software; and a Zeiss 710 confocal microscope with Zeiss LSM Image Browser (version 4.2.0.121) software, which was used to create three-dimensional images and stack movies. Images of sections were taken using a Zeiss AxioSkop 2 MOT microscope fitted with a QImaging Retiga 2000R camera, a Qimaging RGB pancake and QCapture Pro 6.0 software; a Zeiss Scope.A1 microscope fitted with a Zeiss AxioCam MRm camera and Zeiss ZEN 2012 (blue edition) software; and a Zeiss LSM 780 confocal microscope with Zeiss ZEN 2011 (black edition) software. All images were further processed in Photoshop CS4 (Adobe Systems Inc., San Jose, CA) and/or ImageJ 1.50i software (NIH, Bethesda, MD).

Hockman D., Burns A.J., Schlosser G., Gates K.P., Jevans B., Mongera A., Fisher S., Unlu G., Knapik E.W., Kaufman C.K., Mosimann C., Zon L.I., Lancman J.J., Dong P.D., Lickert H., Tucker A.S, & Baker C.V. (2017). Evolution of the hypoxia-sensitive cells involved in amniote respiratory reflexes. eLife, 6, e21231.

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Histological Analysis of Plant Tissues

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The material collected was processed according to Travaglia et al. (2012) . Micro-sections (10 μm) of leaf midrib and berry pedicels (at the berry proximal zone) were prepared using a rotary microtome, whereas stem cross-sections were prepared by hand-cut. The histological preparations were photographed with a camera (AxioCam HRc camera, Carl Zeiss, Göttingen, Germany) attached to a standard microscope (Model 16, Carl Zeiss) . The boundary between xylem and phloem was easily identified because of the differential staining of the cell walls corresponding to sieve and vessel elements. The tissue area was calculated using the software Image Pro-Plus (Media Cybernetics Inc, Rockville, MD). Stomata density was assessed by taking two imprints of the abaxial surface of a fully expanded leaf (13th from the shoot apex) with transparent nail varnish. Imprints were performed in middle zone between the main midrib and the leaf margin. When imprints dried, they were mounted onto a slide for examination under optical microscope (40×). Three representative photographs per imprint were taken using an AxioCam HRc camera attached to a Zeiss Axiophot (Carl Zeiss) microscope. Stomata density was calculated as the mean value of the number of stomata per square millimeter of six photographs per leaf. The Image Pro-Plus software (Media Cybernetics Inc) was used to analyze the microphotographs.

Murcia G., Pontin M., Reinoso H., Baraldi R., Bertazza G., Gómez-Talquenca S., Bottini R, & Piccoli P.N. (2016). ABA and GA3 increase carbon allocation in different organs of grapevine plants by inducing accumulation of non-structural carbohydrates in leaves, enhancement of phloem area and expression of sugar transporters. Physiologia plantarum, 156(3).

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Imaging Protocols for BABB-cleared and Glycerol-cleared Embryos

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Images of BABB-cleared embryos were taken using a Zeiss Axiovert 200 microscope with an AxioCam HRc camera and AxioVision Rel 4.8.2 software. Glycerol-cleared embryo images were taken using a Leica MZ APO microscope with a Lumenera Infinity 3 camera and Infinity Analyze software. Sections were imaged using a Zeiss Axiovert 200 microscope with a Lumenera Infinity 3 camera and Micro-Manager 1.4.22 software. Images were cropped and altered for brightness and contrast using Adobe Photoshop CS5.1.

Parker H.J., Bronner M.E, & Krumlauf R. (2019). An atlas of anterior hox gene expression in the embryonic sea lamprey head: hox-code evolution in vertebrates. Developmental biology, 453(1), 19-33.

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Immunohistochemical Analysis of Splenic B Cells

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Spleens from NP-Ficoll and NP-LPS immunized mice were harvested on day 7 and NP-KLH immunized mice on day 10, embedded in OCT, frozen in liquid nitrogen cooled 2-methyl-butane, sectioned, and fixed with acetone. Spleen sections were stained with B220-biotin (BioLegend) and anti-Igλ-AP (Southern Biotech). Streptavidin-HRP (Southern Biotech) was used as a secondary antibody. AP and HRP were detected using Fast-Blue BB base and 3-amino-9-ethylcarbazole (Sigma), respectively. Sections were imaged using a Zeiss Axioplan microscope with a Zeiss Plan-Apochromat 10x/0.32 objective and Zeiss AxioCam HRC camera using AxioVision 4.7.1 software. The images were then processed using Adobe Photoshop CC software.

Merlo L.M., DuHadaway J.B., Montgomery J.D., Peng W.D., Murray P.J., Prendergast G.C., Caton A.J., Muller A.J, & Mandik-Nayak L. (2020). Differential Roles of IDO1 and IDO2 in T and B Cell Inflammatory Immune Responses. Frontiers in Immunology, 11, 1861.

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Quantitative Analysis of Mitochondrial Morphology

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Free-floating coronal sections were incubated in a mixture of primary antisera (Table 1) in phosphate-buffered saline (PBS) containing 0.3% Triton X-100 overnight at room temperature and subsequently reacted with a mixture of FITC- and Cy3-conjugated IgG (or streptavidin, Jackson Immunoresearch Laboratories Inc., West Grove, PA, USA). Negative controls were obtained by omitting primary antibody to verify the specificity of the antibodies. Images were captured using an Axiocam HRc camera and AxioVision Rel. 4.8 Software or a confocal laser scanning microscope (LSM 510 META, Carl Zeiss Inc, Oberkocken, Germany). Fluorescent intensity was measured, and standardized by setting the threshold level (mean background intensity obtained from five image input). Manipulation of the images was restricted to threshold and brightness adjustments to the whole image. Individual mitochondrion length in CA1 pyramidal cells (n = 20/section) was also measured with a 100× objective lens by AxioVision Rel. 4.8 Software.

Hyun H.W., Ko A.R, & Kang T.C. (2016). Mitochondrial Translocation of High Mobility Group Box 1 Facilitates LIM Kinase 2-Mediated Programmed Necrotic Neuronal Death. Frontiers in Cellular Neuroscience, 10, 99.

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Neutral Red Staining of Microglia

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Neutral red was dissolved in distilled water to give a stock solution (2.5 mg/ml), which is stable at room temperature for several months. Staining using neutral red was done by treating larvae at 4 dpf with solution (5 μg/ml) of neutral red in embryo water containing PTU for 3 hours (46 (link)). Animals were rinsed at least twice after neutral red staining and washed overnight in embryo water with PTU at 28.5°C. Approximately 24 hours after neutral red treatment, larvae were anesthetized and mounted in 1.5% low–melting point agarose. The number of neutral red⁺ microglia was counted, and images were acquired immediately after counting using a Zeiss AxioCam HRc camera with the AxioVision software. Between 100 and 150 embryos were mounted and imaged for each double, triple, and quadruple mutant experiment. After imaging, all animals were genotyped by PCR and statistical analysis was performed.

Iyer H., Shen K., Meireles A.M, & Talbot W.S. (2022). A lysosomal regulatory circuit essential for the development and function of microglia. Science Advances, 8(35), eabp8321.

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Whole-Mount In Situ Hybridization Protocol

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Digoxygenin- and fluorescein-labelled probes were generated by standard methods and used in single and double lamprey whole-mount in situ hybridisation as described previously^{19 (link)}. Embryos were cleared in a solution of 75% glycerol prior to being imaged using a Leica MZ APO microscope with a Zeiss Axiocam HRc camera and Axiovision Rel 4.8 software. Images were cropped and altered for brightness and contrast using Adobe Photoshop CS5.1.

Parker H.J., Bronner M.E, & Krumlauf R. (2014). A Hox regulatory network of hindbrain segmentation is conserved to the base of vertebrates. Nature, 514(7523), 490-493.

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Imaging methods for zebrafish embryos

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All images were acquired at room temperature. Bright field images of live embryos mounted in 3% methyl cellulose/1× tricaine (Sigma-Aldrich) in embryo medium were acquired using a Leica M165FC stereo microscope with DFC425C camera and Leica Application Suite V3.8 software. Images of DAB-stained embryos mounted in benzyl alcohol/benzyl benzoate were obtained with a Zeiss Axiophot microscope, 20× Plan-Neofluar objective, AxioCam HRc camera, and Axiovision software. Confocal images of embryos mounted in low-melting-point agarose in E3 embryo medium were obtained using a Zeiss LSM 710 or LSM 700 confocal microscope, 40×/1.1 W Korr LD C-Apochromat, or 20×/0.8 Plan-Apochromat objective and Zen 2.3 SP1 software. Images were processed using Fiji/ImageJ software, including generation of orthogonal projections, maximum intensity projections, and adjustment of brightness and contrast.

Armistead J., Hatzold J., van Roye A., Fahle E, & Hammerschmidt M. (2019). Entosis and apical cell extrusion constitute a tumor-suppressive mechanism downstream of Matriptase. The Journal of Cell Biology, 219(2), e201905190.

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Whole-Mount In Situ Hybridization Protocol

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Parker H.J., Bronner M.E, & Krumlauf R. (2014). A Hox regulatory network of hindbrain segmentation is conserved to the base of vertebrates. Nature, 514(7523), 490-493.

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Lamprey embryo in situ hybridization

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Digoxygenin-labelled probes were generated by standard methods and purified using the MEGAclear Transcription Clean-Up Kit (Ambion). Lamprey embryos were staged according to Tahara et al.⁶⁹. Lamprey whole-mount in situ hybridisation was performed on MEMFA-fixed embryos following established protocols^{70 (link)}, with the following additions to the protocol:^{71 (link)} methanol-stored embryos were transferred into ethanol and left overnight prior to rehydration; embryos were treated with 0.5% acetic anhydride in 0.1 M triethanolamine after proteinase K treatment. For imaging, embryos were cleared in benzyl alcohol:benzyl benzoate and mounted in Permount (Fisher Scientific).
For sectioning after in situ hybridisation, embryos were transferred into 30% sucrose in phosphate-buffered saline, embedded in O.C.T. Compound and sectioned to 10-µm-thick cryosections. Images were taken using a Zeiss Axiovert 200 microscope with AxioCam HRc camera and AxioVision Rel 4.8.2 software.

Parker H.J., De Kumar B., Green S.A., Prummel K.D., Hess C., Kaufman C.K., Mosimann C., Wiedemann L.M., Bronner M.E, & Krumlauf R. (2019). A Hox-TALE regulatory circuit for neural crest patterning is conserved across vertebrates. Nature Communications, 10, 1189.

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