Fifteen minutes after the start of reperfusion, left ventricular tissue was prepared for cryosectioning by freezing in Tissue‐Tek OCT Compound (Sakura Finetek Germany GmbH, Staufen, Germany). To perform dihydroethidium staining, cryoslices of the LV were incubated with the fluorescent probe dihydroethidium (dissolved in 1× PBS) for 10 minutes at 37°C in a light‐protected humidity chamber, then fixed with Dako Fluorescent Mounting Medium (Dako North America Inc, Carpinteria, CA). Sections were imaged by fluorescence microscopy (LSM 510 META; Carl Zeiss, Jena, Germany) using an excitation wavelength of 488 nm; emission was recorded at 540 nm.19 Analysis was performed by digital image analysis using Leica Confocal Software Lite Version (LCS Lite; Leica, Wetzlar, Germany). The mean fluorescence intensity of n=6 ventricles was used to quantify the extent of superoxide.
Confocal software lite version lcs lite
Manufactured by Leica
Sourced in Germany
Leica Confocal Software Lite Version (LCS Lite) is a software application designed for basic confocal microscopy image acquisition and processing. It provides essential functions for controlling the confocal microscope and capturing high-quality digital images.
Automatically generated - may contain errors
3 protocols using confocal software lite version lcs lite
1
Superoxide Imaging in Cardiac Reperfusion
2
Quantifying Interstitial Collagen Fibrosis
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Samples were embedded with Tissue-Tek® (Sakura, Alphen, Netherlands) and sectioned in 10 μm slices. Histological sections were fixed in Bouin solution and subsequently stained in 0.1% (wt/vol) Sirius red solution (Sigma-Aldrich Chemie, Steinheim, Germany). Sections were washed by 0.01 N HCl, Aqua dest. and counterstained for nuclei by Mayers hemalaun solution, washed with Aqua dest. for 5 min and dehydrated with ethanol. Finally, histological slices were visualized under light microscopy. Total collagen content was quantified by digital image analysis using Leica Confocal Software Lite Version (LCS Lite). The mean of n = 6 preparations was used to quantify the extent of interstitial fibrosis.
3
Quantifying Superoxide Levels in Cardiac Tissue
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To perform DHE staining, cryosections of the left ventricle (LV) were incubated with DHE (dissolved in 1 X PBS) for 10 min at 37°C in a light-protected humidity chamber, then fixed with Dako Fluorescent Mounting Medium (Dako, North America Inc., USA). Slides were imaged by fluorescence microscopy (LSM 510 META; Carl Zeiss, Jena, Germany) using an excitation wavelength of 488 nm; emission was recorded at 540 nm (Nazarewicz et al., 2013 (link)). Analysis was performed by digital image analysis using Leica Confocal Software Lite Version (LCS Lite). The mean fluorescence intensity of n = 4 ventricles was used to quantify the extent of .
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