The mesenteric arterial bed was perfused with Bouin’s solution to fix the mesenteric arteries. Second order arteries were isolated, permeabilized with 0.01% Triton X-100 and blocked with 3% BSA, as described previously67 (link). Vessels were incubated overnight at 4 °C with an anti-CGRP primary antibody (Thermo Scientific, Rockford, IL, USA), and then, with an Alexa-568-labeled goat anti-mouse secondary antibody (Molecular Probes, Eugene, OR, USA) for 4 h at 4 °C. The fluorescent signal was examined with an Olympus BX41 WI microscope and a CCD camera (Jenoptik ProgRes C5).
Progres c5
Manufactured by Jenoptik
Sourced in Germany
The ProgRes C5 is a high-resolution digital camera designed for microscopy applications. It features a 5-megapixel CMOS sensor and captures images with a resolution of 2560 x 1920 pixels. The camera connects to a computer via a USB interface and is compatible with various microscopy software solutions.
Automatically generated - may contain errors
Lab products found in correlation
Bx41 wi microscope, by Jenoptik (2 mentions)
Signal enhancer hikari, by Nacalai Tesque (1 mentions)
Cytoseal, by Thermo Fisher Scientific (1 mentions)
Dmire2, by Leica (1 mentions)
Progres mac capturepro 2, by Jenoptik (1 mentions)
Isoproterenol, by Merck Group (1 mentions)
Feinoptic microscope, by Jenoptik (1 mentions)
Microtome, by Leica (1 mentions)
Axioplan microscope, by Jenoptik (1 mentions)
Axiostar plus, by Zeiss (1 mentions)
Mayer s hemalum solution, by Merck Group (1 mentions)
12 protocols using progres c5
1
Mesenteric Artery Immunofluorescence Staining
2
Immunofluorescence Staining of Astrocytes
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Astrocyte monolayers were fixed with 2% paraformaldehyde in PBS, blocked with 0.5% BSA in PBS and incubated with the corresponding rabbit polyclonal primary antibody, mouse monoclonal primary antibody or both in the case of co-immunofluorescence analysis, and then, with the appropriate Alexa-568-labeled goat anti-rabbit or anti-mouse secondary antibody (Invitrogen Molecular Probes, USA, Cat. #A11011 or Cat. #A11004, respectively), or Alexa-488-labeled goat anti-rabbit or anti-mouse secondary antibody (Invitrogen Molecular Probes, USA, Cat. #A11008 or Cat. #A10680, respectively) using the Signal Enhancer HIKARI (Nacalai Tesque, INC, Japan) as indicated by the manufacturer. The fluorescent signal was examined using an Olympus BX41 WI microscope and a CCD camera (Jenoptik ProgRes C5).
3
Microstructural Analysis of Muscle-Capsule Interface
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The microstructural characteristics of the muscular fibres attaching onto the joint capsule were studied histologically. Seven specimens were randomly selected from the total of 20 dissected upper limbs, where the muscular fibres were excised together with a portion of the adjacent triceps brachii muscle and surrounding joint capsule at the point of their insertion. These en bloc harvested specimens were then cut to form a 1 × 1 × 1 cm cube with the central area representing the close relationship between the capsule and the muscular fibres. Subsequently, the collected samples were fixed in a 4% formaldehyde and sent for embedding. Each specimen underwent dehydration in an increasing series of alcohol, clearing in a xylene bath, infiltration with melted paraffin, and embedding in a paraffin wax. Parallel sections were cut at 4 μm, and stained with haematoxylin–eosin. All specimens were assessed using light microscope BX53 (Olympus) with a CCD camera ProgRes C5 (Jenoptik). The digital images were processed through the image analysis software NIS‐Elements (Laboratory Imaging). Two skilled histologists observed all specimens and interpreted the findings (T.N. and J.U.).
4
Syncytial Contractility under Hydrostatic Pressure
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On day 3 of culture, at a constant temperature of 37°C and 5% CO2, each syncytium was observed via a movie capture system (ProgRes C5, Jenoptik, Germany) in four different conditions of relative hydrostatic pressure: at 0 (atmospheric pressure), 100, 200, and 300 mmHg inside a custom-machined polymethylmethacrylate pressure chamber (Figure 1 ).
The syncytium cultures were repeated 4 times for a total of 40 syncytia loaded by both increasing and decreasing pressures. In particular, for each syncytium, we have obtained a sequence of eight video files in AVI format, each video with a duration of 40 s: at 0, 100, 200, and 300 mmHg (after that there was a recovery of 0.5 h in incubator at 37°C and 5% CO2) and then at 300, 200, 100, and 0 mmHg.
The syncytium cultures were repeated 4 times for a total of 40 syncytia loaded by both increasing and decreasing pressures. In particular, for each syncytium, we have obtained a sequence of eight video files in AVI format, each video with a duration of 40 s: at 0, 100, 200, and 300 mmHg (after that there was a recovery of 0.5 h in incubator at 37°C and 5% CO2) and then at 300, 200, 100, and 0 mmHg.
5
Histological Analysis of Mammary Gland
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Paraffin-embedded mammary gland sections (6 µm) were deparaffinized in xylene for 10 min, rehydrated in descending grades of ethanol baths, and stained with 1% Harris's haematoxylin and 1% eosin. Sections were dehydrated in ascending grades of ethanol and xylene baths and mounted with Cytoseal (Richard-Allan Scientific). Qualitative histological analysis was performed by imaging several arbitrary areas per 20× field of view using a Leica DM IRE2 inverted epifluorescence microscope equipped with a ProgRes C5 camera (Jenoptik) and ProgRes Mac CapturePro 2.7.6 imaging software (N = 8).
6
Cardiac Syncytium Contractility Dynamics
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On day 3 of culture, at a constant temperature of 37°C and 5% CO2, each syncytium was observed via a movie capture system (ProgRes C5, Jenoptik, Germany) in four different conditions: untreated control (CTRL); stimulus via β-adrenergic isoproterenol (ISO, 10 μM; Sigma-Aldrich, Milan, Italy); stimulus via an electromagnetic field (EMF; see below for details); and stimulus via both isoproterenol and electromagnetic field (ISO + EMF). In particular, for each condition, AVI videos (duration, 20 s) of 20 beating syncytia were collected every 3 min, permitting us to specifically study the average contraction pattern during the time interval 27–39 min.
7
Quantifying Pancreatic Tissue Changes
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Pancreatic tissue specimens from Kras and Kras/Slug mice were stained for amylase, CK19, PCNA, p-MLC2(S19) and p-ERK1/2(T202/Y204). Antigen retrieval was conducted as previously described10 (link)13 (link). Photographs for quantitative comparison were taken using FeinOptic microscope and Jenoptik ProgRes C5 camera. Number of Kras and Kras/Slug mice with no staining (0), less than 5% (1+), 5% to 25% (2+), or more than 25% (3+) of the pancreas with CK19 staining was determined. Relative number of p-ERK1/2(+) and PCNA(+) nuclei in Kras and Kras/Slug mice were quantified.
8
Sectioning and Staining Mouse Embryos
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E10.5, E11.5, and E14.5 embryos were fixed in 4% PFA in PBS at 4°C, rinsed in PBS, dehydrated through an ethanol series, and then embedded in paraffin. Six-micron sections were cut using a Leica microtome, mounted on slides, and stained with Harris modified hematoxylin and Eosin Y using a standard protocol. Images were taken using a Zeiss Axioplan microscope fitted with a Jenoptik ProgRes C5 camera. If needed, images were merged using the Adobe Photoshop Photomerge function.
9
Quantifying Osteoblastic Mineralization in Cell Cultures
10
Quantifying Micronuclei and Binucleation in E. carteri
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Relative abundance of micronucleated and binucleated cells of E. carteri was estimated after the modified protocol of Fenech et al. (2003 (link)) and Bolognesi and Fenech (2012 (link)). The Giemsa stained cells were examined under light microscope (Axiostar Plus; Zeiss Microscopy, Jena, Germany) for different kinds of nuclear aberrations. Digital photodocumentation and analyses of different nuclear anomalies of E. carteri cells was carried out using a CCD camera (ProgRes C5; Jenoptik, Jena, Germany) attached to the light microscope (BH-2; Olympus, Tokyo, Japan). Round or ovoid shaped non-refractory particles with the color and structure identical to chromatin with diameter equaling 1/3–1/20 of the main nucleus and its clear detachment from macronucleus were interpreted as ‘micronuclei’ (Chakraborty & Ray, 2009 ). On the other hand, binucleated cells consisted of two nuclei with intact nuclear membrane and identical staining intensity and were found within the same cytoplasmic boundaries. At least 450 cells were examined per slide for the enumeration of micronucleated and binucleated cells in E. carteri treated with washing soda.
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