Pictures of adrenal glands (H&E staining and in situ hybridization) and hearts (Sirius red staining) were recorded with an Axioplan 2 microscope equipped with a Hal100 microscope lamp and an Axiocam MRc5 camera using Plan-Neofluar (5×, 10×, 40×) objectives and Axiovision Rel 4.8 software (all from Carl Zeiss). Pictures of whole kidneys (H&E staining) were recorded with an Axioscop 40 equipped with an Axiocam MRc5 and MCX-2eco high-resolution position controller using a Plan-Apochromat 20× objective and Axiovision Rel 4.8 software (all from Carl Zeiss).
Axiocam mrc5
Manufactured by Zeiss
Sourced in Germany, United States, Italy, Switzerland, Netherlands
The AxioCam MRc5 is a high-performance digital camera designed for microscopy applications. It features a 5-megapixel CMOS sensor that captures images with a resolution of 2584 x 1936 pixels. The camera is capable of delivering fast live image preview and recording at up to 30 frames per second. It is compatible with a variety of microscope systems and supports multiple image file formats, including TIFF, JPEG, and BMP.
Automatically generated - may contain errors
Lab products found in correlation
Axiovert 200m, by Zeiss (33 mentions)
Axioplan light microscope, by Zeiss (27 mentions)
Axioplan microscope, by Zeiss (22 mentions)
Axiovision software, by Zeiss (20 mentions)
Axiovision 4, by Zeiss (18 mentions)
Axio observer z1, by Zeiss (14 mentions)
Axiocam mrm, by Zeiss (12 mentions)
Axio imager, by Zeiss (12 mentions)
Axiovision rel 4, by Zeiss (11 mentions)
Axioskop 40, by Zeiss (10 mentions)
Axioplan 2, by Zeiss (9 mentions)
Axiovision release 4 8 2 sp2, by Zeiss (9 mentions)
Axio imager a1 microscope, by Zeiss (9 mentions)
Axiovision se64, by Zeiss (9 mentions)
Axiovision v4, by Zeiss (9 mentions)
Axio observer a1, by Zeiss (9 mentions)
Axio imager z2, by Zeiss (8 mentions)
Axio imager a1, by Zeiss (8 mentions)
Axiovert 200m microscope, by Zeiss (8 mentions)
Zen software, by Zeiss (8 mentions)
573 protocols using axiocam mrc5
1
Imaging of Adrenal Glands, Hearts, and Kidneys
2
Techniques for Visualizing Embryo Development
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To visualize electroporation targeting or in situ hybridization results, whole-mount embryos were imaged in PBS + 0.1% Tween using a stereoscope (Discovery V8; Carl Zeiss) with an Achromat S 1.0× lens and fluorescence module outfitted with a GFP 500 filter cube. Images were acquired with a digital camera (AxioCam MRc5; Carl Zeiss) using Axiovision 4.8.2 software (Carl Zeiss). Slides and cultures were mounted with Permafluor (Thermo Fisher Scientific) containing 1 mg/ml DAPI. In situ hybridized sections were imaged on a microscope (Axioimager A1; Carl Zeiss) with a Plan Apochromat 20×/0.8 NA lens and acquired with the AxioCam MRc5 camera using Axiovision 4.8.2 software. Immunofluorescently labeled cells and sections were imaged using a confocal system (LSM 710; Carl Zeiss) attached to an inverted microscope (Observer Z.1; Carl Zeiss) outfitted with Plan Apochromat 10×/0.45 NA, 20×/0.8 NA, 40×/0.95 NA, and 100×/1.46 NA lenses and FS38, FS43, and FS49 filter sets. Confocal images were acquired using ZEN 2010 software (Carl Zeiss), and two-dimensional maximum intensity projections were produced from three-dimensional z-stack images using the data from the highest intensity pixels along the projection axis. All imaging was performed at room temperature. Image files were assembled in Photoshop (Adobe).
3
Quantifying Fluorescence Intensity in PDLCs
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After staining and mounting, the stained slides were separated from the supports and images were acquired with a fluorescence microscope equipped with an ApoTome slide module and AxioVision 4.8 software, using 40×/1.0 objective lens (Zeiss AxioCam MRc5; Carl Zeiss Microimaging). The images were examined and photographed under the same acquisition hue, saturation, and intensity (HSI) threshold settings. The fluorescence intensity of secondary antibody was measured using Fiji 1.51n in‐built software on fluorescence micrographs of PDLCs obtained without DAPI nuclear staining. The relative fluorescence intensity compared with control was quantified for each condition under identical threshold settings of color, saturation, and brightness (n = 3) (Cuijpers et al.., 2015 ).
4
Quantifying Neurons in Micrographs Using ImageJ
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Using ImageJ software (version 1.53k), we counted every labeled neuron visible in high-resolution micrographs of all the sections containing the NLL. Micrographs were taken with a Zeiss Axioskop 40 microscope equipped with a Zeiss AxioCam MRc 5 digital camera (Carl Zeiss, Oberkochen, Germany). To correct for oversampling due to split neurons appearing in two adjacent sections, we applied Abercrombie’s formula (Abercrombie, 1946 (link)):
where N is the estimated number of labeled neurons, n is the number of neurons counted, T is the section thickness, and D is the mean maximum diameter of the labeled cell bodies. To calculate the mean cell body diameter, we used the case with the most labeled neurons (case FG01) and measured the maximum diameter of every labeled neuron with sharp cell body contour found in each ipsilateral NLL (seeTable 1 below). To measure the maximum diameter, we traced the contour of the cell body on high-resolution micrographs using the polygon tool of ImageJ.
where N is the estimated number of labeled neurons, n is the number of neurons counted, T is the section thickness, and D is the mean maximum diameter of the labeled cell bodies. To calculate the mean cell body diameter, we used the case with the most labeled neurons (case FG01) and measured the maximum diameter of every labeled neuron with sharp cell body contour found in each ipsilateral NLL (see
5
Immunostaining of C3aR and C3 in Embryos
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After an overnight fixation in 4% PFA, embryos were washed with 1% PBST (1% Triton-X100 in PBS) and incubated for 2 h in blocking solution (10% goat serum, 0.2% sodium azide in 1% PBST). C3aR or C3 Abs (1:100 in blocking solution) were added and incubated at 4 °C for three days following several washes with 10% goat serum in 1% PBST and 1% PBST only. A solution of secondary antibody (1:100 in 10% goat serum in 1% PBST) was added to the embryo and incubated at 4 °C for 2 days followed by several washes with 1% PBST and PBS. Embryos were analyzed using a SteREO Discovery V12 microscope (Carl Zeiss, Göttingen, Germany) and the images were captured using Zeiss AxioCam MRc5 with AxioPlan 2 and Zeiss AxioVison 4.7.1 extended focus module (AxioVision, Göttingen, Germany). A blocking peptide or pre-immune IgG was used as a negative control to show non-specific binding of C3aR or C3 Abs as previously described (Haynes et al., 2013 (link)). The results represent at least three different biological replicates.
6
Histological Analysis of PM2.5-Induced Lung Damage
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Lungs from sham and PM2.5win-treated mice were properly inflated, excised and immediately formalin fixed and processed for routine histology. Briefly, after being preserved for 24 h in the fixative, tissue samples were rinsed in distilled water, dehydrated in an ethanol series from 70% to 100% and embedded in Bio-plast tissue embedding medium. For each sham and PM2.5win exposed lung sample, 7 µm serial sections were cut by a rotary microtome, mounted on slides and stained with Mayer's haemalaun and alcoholic eosin. Samples were qualitatively screened by means of Zeiss Axioplan microscope at a magnification of 40× and images were taken using Zeiss AxioCam MRc5 digital camera interfaced with the Axiovision Real 4.6 software. Figure panels were prepared by means of Adobe Photoshop.
7
Morphological Analysis of Cultured Fungal Strains
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Morphological and anatomical characters of the cultured strains were analyzed using light microscopy and documented with digital photographs as in Muggia et al. [9 (link)]. Analyses and photographs were performed on 10 month to one year old subcultures considering the following characters: form of growth, branching of the hyphae and melanization. Small fragments of the mycelia were taken; squashed sections were mounted in water and studied by light microscopy. All images were acquired with a ZEISSAxioCam MRc5 (Zeiss, Jena, Germany) digital camera fitted to the microscope. Both images of growth habit and hyphae structure [9 (link)] were digitally processed using the CombineZM software [53 ]. The photos were slightly refined in sharpness and color tone with Adobe Photoshop 7.0 (© Adobe System Incorporated, San Jose, CA, USA) and the figures were prepared with CorelDRAW X7 (© Corel Corporation, Ottawa, Canada).
8
Fixation and Phosphatase Staining
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Samples were washed
twice with PBS and fixed with 4% PFA. After fixation, the samples
were washed twice with PBS. The ELF97 dye was diluted 50 times using
the above acid phosphatase solution, added to each well, and incubated
for 15 min at 37 °C in the dark. The samples were then washed
twice with PBS, and images were taken using a microscope (Zeiss AxioCam
MRc5; Carl Zeiss Microimaging, Germany).
twice with PBS and fixed with 4% PFA. After fixation, the samples
were washed twice with PBS. The ELF97 dye was diluted 50 times using
the above acid phosphatase solution, added to each well, and incubated
for 15 min at 37 °C in the dark. The samples were then washed
twice with PBS, and images were taken using a microscope (Zeiss AxioCam
MRc5; Carl Zeiss Microimaging, Germany).
9
Immunostaining of C3aR and C3 in Embryos
10
Tissue Preparation and Histological Analysis
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All samples were washed in Phosphate Buffered Saline (PBS; Sigma, Milan, Italy) and subsequently were fixed in 10% buffered formalin for 2 h. The dehydration and clarification process were carried out the following day using graduated ethanol and xylene. Finally, the tissues were paraffin-embedded without altering their anatomical orientation, as previously described [23 ]. Using a microtome, 4–5 μm thick sections were obtained, which were maintained at room temperature after being mounted on silane-coated slides (Dako, Glostrup, Denmark). All sections were observed for general morphological analysis with a Zeiss Axioplan light microscope (Carl Zeiss, Oberkochen, Germany) using Hematoxylin and Eosin (H&E) staining. Best representative photomicrographs were captured using a Zeiss AxioCam MRc5 digital camera (Carl Zeiss). Two independent pathologists have analyzed the slides.
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