Fluorescent images were taken using the confocal microscopes Leica TCS SP5 and Leica TCS SP-II. Image analysis was performed using the Fiji ImageJ software and Adobe Photoshop. To quantify the percentage of Col XII in the post injured myocardium, the area positive for Col XII expression was divided by the whole post-injury myocardium area, but excluding the epicardium. N represents a number of hearts. In order to obtain representative data, at least 2 sections of each heart at different time points were imaged and analyzed. Statistics were calculated with the Prism GraphPad software. All results are expressed as the mean ± standard error of the mean (S.E.M.). Bright-field images were taken using a Zeiss Axioplan 2 microscope coupled to an AxioCam Color camera.
Axiocam color camera
Manufactured by Zeiss
Sourced in Germany
The Axiocam color camera is a high-performance imaging device designed for use in microscopy applications. It features a color image sensor and delivers detailed, high-resolution images for various scientific and industrial purposes.
Automatically generated - may contain errors
Lab products found in correlation
Axioplan 2 microscope, by Zeiss (4 mentions)
Axiovision software, by Zeiss (2 mentions)
Axiovision 4, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Tcs spii, by Leica (1 mentions)
Tcs sp5, by Leica (1 mentions)
Image j software, by Adobe (1 mentions)
Stereomicroscope, by Zeiss (1 mentions)
Bm purple, by Roche (1 mentions)
Rna labeling kit, by Roche (1 mentions)
Axioplan microscope, by Zeiss (1 mentions)
Rabbit anti laminin antibody, by Merck Group (1 mentions)
Congo red, by Merck Group (1 mentions)
Axiovert 200, by Zeiss (1 mentions)
Cm1900 cryostat, by Leica (1 mentions)
Ncl p53 cm5p, by Leica (1 mentions)
Axio imager a1 microscope, by Zeiss (1 mentions)
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
Frozen section medium, by Thermo Fisher Scientific (1 mentions)
Rabbit anti human ige, by Agilent Technologies (1 mentions)
16 protocols using axiocam color camera
1
Quantifying Collagen XII in Post-Injury Myocardium
2
Whole Mount In Situ Hybridization Protocol
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Whole mount in situ hybridization was performed as previously described51 (link) using an automated InsituPro system (Abimed, Langenfeld, Germany).52 Digoxigenin-labeled RNA probes were prepared using an RNA labeling kit (Roche, Indianapolis, IN, USA) and stained using BM purple (Roche). Whole mount embryos were imaged on a Leica stereomicroscope fitted with a Zeiss Axiocam color camera. For generating in situ probes, following PCR primers were used (forward and reverse primers are flanked with T3 and T7 promoter sequences respectively): zfFam40b-Forw-T3–5′-AATTAACCCTCACTAAAGGGAATGATACAGATAC-3′ zfFam40b-Rev-T7–5′-TAATACGACTCACTATAGGGTTGCTCTGGTTCTT-3′ vmhc-Forw-T3–5′-AATTAACCCTCACTAAAGGGAGGAAAGAGCATCC-3′ vmhc-Rev-T7-5′-TAATACGACTCACTATAGGGCCCACATCACGACT-3′ amhc-Forw-T3–5′-AATTAACCCTCACTAAAGGGCGAGATATGAGACT-3′ amhc-Rev-T7-5′-TAATACGACTCACTATAGGGTTTCTTTCAAGCAT-3′ cmlc2-Forw-T3–5′-AATTAACCCTCACTAAAGGGATGGAGTTATCAAC-3′ cmlc2-Rev-T7-5′-TAATACGACTCACTATAGGGCAGCAGTTACAGAC-3′.
3
Embryonic Vascular Morphometry Analysis
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Serial transverse sections (4 μm) of formalin-fixed E13.5 embryos were cut using a microtome. Slides were dried overnight at 37 °C and heated for 20 min at 60 °C. Haematoxylin and eosin staining of every tenth section was used to select the anatomical level according to the ref. 54 . IHC for CD31, NG2 and DOCK4 was carried out using standard methods. Images were captured using an Axioplan Zeiss microscope fitted with AxioCam color camera and AxioVision 4.6 software (Carl Zeiss). Images at the brain level were captured using a × 20 objective and at the lung level using a × 4 objective. Vessel diameter for each lumen was defined, and the best fit circle for each lumen was determined using the AxioVision software (smallest identifiable lumens were 8 μm).
4
Quantitative Analysis of Cerebral Amyloid Angiopathy
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Mice were saline/heparin-perfused, and 20-µm coronal brain cryostat sections were fixed with 4% paraformaldehyde. Brain sections were incubated with rabbit anti–laminin antibody (Sigma-Aldrich) overnight and stained for 30 min with 0.2% Congo Red (Sigma) in 70% isopropanol. After immunohistochemistry, brain sections were analyzed with a microscope (Axiovert 200; Carl Zeiss) equipped with Plan-Neofluar (10× NA 0.3, and 20× NA 0.5) objective lenses at room temperature. The imaging medium was air for both the objective lenses used. The AxioCam color camera (Carl Zeiss) and AxioVision software (Carl Zeiss) were used for image collection. Each set of stained sections was processed under identical gain and laser power setting and under identical brightness and contrast settings. Images of all the areas with CAA and Aβ plaques were acquired and thresholded using Image J. The total area of CAA and Aβ plaques was analyzed as percentage of total cortex area with the analyzer blinded to treatment of mice. The average of 7–10 different sections from each mouse was determined (n = 5 mice per group).
5
Decalcification and Cryosectioning of Bone Samples
6
Immunohistochemical Analysis of Mouse Organs
7
Immunofluorescence Staining of Tumor Cryosections
8
Cardiomyocyte Nucleation Analysis
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Serial sections (0.5–1 μm) were cut from LR white embedded heart to cover up to 100 μm of tissue below and above the section that was mounted on the chip. Sections were stained with a modified PAS staining protocol to identify individual cardiomyocytes. Slides were immersed for 2 × 30 min in xylene and rehydrated through graded alcohols, incubated in Periodic acid solution overnight and then in Schiff’s reagent for two nights, washed, and dehydrated and cleared before mounting. Slides were imaged on a NanoZoomer Whole Slide Scanner (Hamamatsu Photonics). Magnified PAS image in Fig. 3a was taken on Airy Scan LSM800 with Axiocam color camera (Zeiss). To analyze nucleation, 15N-thymidine-positive cardiomyocytes were tracked by locating the cardiomyocyte in every section of the serial above and below the level of the nucleus for as long as it was present. The total number of nuclei was counted for each cell. PAS-stained sections were also used to confirm cells labeled as cardiomyocytes and non-CMs in all cohorts and adjustments have been made for mislabeled cells if necessary. Outlined 15N-positive cells were also used to calculate total cellular volume.
9
Quantifying Hepatic Fibrosis with Sirius Red
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Liver collagen content was determined using sirius red histochemistry as previously described [62 (link)]. Tissue sections were observed using an Axiophot microscope (Carl Zeiss AG, Feldbach, Switzerland) and images were acquired with an Axiocam color camera (Zeiss, Feldbach, Switzerland). Hepatic fibrosis extent was determined using morphometric quantification (MetaMorph Software, Universal Imaging, West Chester, PA, USA).
10
Quantifying Cerebral Infarct Volumes
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