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.
Axioplan microscope
Manufactured by Jenoptik
Sourced in Germany
The Axioplan microscope is a versatile optical microscope designed for a wide range of applications in research and industry. It features a modular construction that allows for customization to meet specific user requirements. The microscope's core function is to provide high-quality, detailed images of samples through the use of various illumination techniques and optical components.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Microtome, by Leica (1 mentions)
Progres c5, by Jenoptik (1 mentions)
Ks 400 imaging system, by Zeiss (1 mentions)
Progres c10, by Jenoptik (1 mentions)
Immpact dab substrate kit, by Vector Laboratories (1 mentions)
Tcs sp8 confocal microscope, by Leica (1 mentions)
Progres c5 camera, by Jenoptik (1 mentions)
3 protocols using axioplan microscope
1
Sectioning and Staining Mouse Embryos
2
Quantifying Muscle Fiber Composition
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For each species, the number of fibers and the area occupied by each type of muscle fibers was quantified by morphometric analysis at ×200 magnification. The procedure was based on a hardware system consisting of a Zeiss Axioplan microscope equipped with a ProgRes C10 plus color camera (Jenoptik, Germany) connected to an IBM‐compatible PC, and using software designed for morphometry and color analysis (KS 400 Imaging system, Carl Zeiss Vision GmbH, München, Germany). This image‐analyzing system discriminated between immunoreactive muscle fibers based on differences in color and contrast. For each image, eight standardized microscopic fields of 97.2 μm2 each were selected at random separately in the dorsal and ventral muscular layers. For each lizard (n = 3 per species), the absolute and relative areas occupied by the three categories of muscle fibers were calculated in the ventral and dorsal parts, as well as in the body and tail parts. Each measure was performed in triplicate on three consecutive histological slides. For each type of muscle fiber, the area of the fibers was divided by the total number of fibers to estimate the mean individual area of each type of fiber in cross sectional view in the dorsal and ventral parts (and also in the body and tail areas).
3
Immunohistochemistry and Immunofluorescence Techniques for Protein Localization
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FFPE tissue sections were deparaffinized and rehydrated through a series of decreasing ethanol washes. Heat induced epitope retrieval was performed using a 20mM sodium citrate pH 6.0 solution in a pressure cooker for 30 minutes followed by pressure release and gradual cooling to 45°C. Sections were washed in PBS, blocked in PBS containing 10% goat serum, incubated with primary antibody overnight at 4°C, then with secondary antibody for one hour at room temperature. For IHC, signal was developed using the ImmPACT DAB Substrate Kit (Vector Laboratories SK-4105). Images were taken using a Zeiss Axioplan microscope fitted with a Jenoptik ProgRes C5 camera. For immunofluorescence, images were taken using a Zeiss LSM780 Confocal microscope. For super-resolution STED microscopy, images were taken using a Leica TCS SP8 Confocal microscope. Colocalization was determined using FIJI ImageJ. Threshold values for each channel were calculated for each set of images. Colocalization values were determined using the Colocalization Threshold plugin of ImageJ. One-way ANOVA with Bonferroni correction for multiple comparisons was used to determine statistical significance.
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