Apoptotic cells were assessed using the Annexin V-FITC Detection Kit (Vazyme, China) according to the protocols of the manufacturer. After pre-cooling, 2–3 mm of flesh under the exocarp was cut into 2 mm × 2 mm × 3 mm cuboids on ice, embedded in optimal cutting temperature (OCT), and placed on a quick-freezing rack. A 25 μm-thick sheet was cut using a freezing microtome (Model: HM525, LEICA, Germany). The cells were washed two times with cold PBS and suspended in the binding buffer. The samples were stained with 10 μl of Annexin V-FITC and 10 μl of PI for 20 min at room temperature (25°C) in the dark. The apoptotic index was immediately determined using a confocal laser scanning microscope (TCS SP5 II, Leica, Germany).
Hm525
Manufactured by Leica
Sourced in Germany
The HM525 is a high-performance microtome designed for precise sectioning of tissue samples. It features a stable and precise cutting mechanism, adjustable specimen feed, and a user-friendly interface. The HM525 is a reliable and versatile instrument suitable for a variety of laboratory applications.
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Lab products found in correlation
4 protocols using hm525
1
Annexin V-FITC Apoptosis Assay
2
Immunohistochemical Staining of Brain Tissue
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Brain tissue was cut coronally into 40 µm sections using a MICROM HM 525 or Leica CM1950 Cryostat. Tissue sections were stored in 96 well plates immersed with 1X TBS and 0.001 % sodium azide solution at 4°C. For IHC, tissue sections were stained via a free-floating staining method outlined in detail previously 4, 18 . Briefly, tissue sections underwent antigen retrieval in 1N HCl, were washed in 1X TBS three times and then blocked and permeabilized for one hour in 1X TBS containing 5% Normal donkey serum (NDS, Jackson ImmunoResearch Laboratories, Lot #161590) and 0.5% Triton x-100. Primary and secondary antibodies used were the same as listed above. Nuclei were stained with DAPI (4'6'-diamidino-2-phenylindole dihydrochloride.
ProLong Gold (Invitrogen, Cat# P36934) anti-fade reagent was used to mount the brain sections onto slides. The brain sections were imaged using an IX83 Olympus Microscope and processed using Cell Sens software. Detailed Orthogonal Projections were processed through Constrained Iterative Deconvolution using Cells Sens Software and assembled using ZEN Lite (Zeiss).
ProLong Gold (Invitrogen, Cat# P36934) anti-fade reagent was used to mount the brain sections onto slides. The brain sections were imaged using an IX83 Olympus Microscope and processed using Cell Sens software. Detailed Orthogonal Projections were processed through Constrained Iterative Deconvolution using Cells Sens Software and assembled using ZEN Lite (Zeiss).
3
Multimodal Tissue Imaging Techniques
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Frozen slicer (Thermo Scientific, model: HM525); Cryogenic tissue slicer (Leica, model: CM1950); Fluorescence quantitative polymerase chain reaction (PCR) instrument (Roche, model: Light Cycler 480); Probe in vivo confocal microscope (Nikon, model: NiE-A1plus); Binocular fluorescence/microscope (Leica, model: DM4000B); Analytical balance (Mettler-Toledo instrument Shanghai Co., Ltd., model: MS204S); Constant temperature water bath (Jintan Hualong Experimental instrument Factory, HH-S8); High-speed cryogenic centrifuge (TOMOS limited company,model:3-18R); Glass slide (Fisher Scientific, catalog #12-550-15).
4
Microscopic Analysis of Fruit Skin
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White fruit (W), green fruit (G), and red fruit (R) of ‘Xinqihong’ sampled at 128 DAFB were washed. W and G comprised pure-colored fruits. The skin of W and G fruits was peeled. The CK was red fruit skin. The skin at the most darkly colored position was sampled to observe and photograph transverse sections of the skin. Sections (10 μm thickness) of the darkest peel were cut with a Microm HM 525 cryostat and observed under a Leica DM2500 fluorescence microscope equipped with a 20× objective lens.
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