Dm2500 optical microscope

Manufactured by Leica

Sourced in Germany, United States, United Kingdom

The DM2500 is an optical microscope designed for laboratory use. It features a high-quality optical system for clear and detailed observations. The core function of the DM2500 is to provide magnified views of small samples or specimens.

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33 protocols using dm2500 optical microscope

Comprehensive Plant and Fruit Analysis

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The plant height, stem diameter, leaf thickness, leaf length, leaf width, number of fruits, and percentage of NF were calculated following the methods of (He et al., 2022 (link)). Leaf chlorophyll content was determined as described by (Zhu et al., 2020 (link)) using a SPAD-502 chlorophyll meter (Konica Minolta Inc., Japan).
The histology of leaves and fruit peel from the NF and MF groups were observed via paraffin section as described by (Zhang et al., 2021 (link)). Briefly, the leaf and fruit peel tissues were cut into 2–3 mm sections and embedded in paraffin. The tissues were then cut into 5 μm slices. Subsequently, the slices were dewaxed using xylene and the tissues were stained using safranine and fast-green followed by neutral gum sealing. Finally, the sections were observed and photographed using a DM2500 optical microscope (Leica Microsystems, USA).

Huang Q., Wang N., Liu J., Liao H., Zeng Z., Hu C., Wei C., Tan S., Liu F., Li G., Huang H., Chen D., Wei S, & Qin Z. (2023). Soil bacterial communities associated with marbled fruit in Citrus reticulata Blanco ‘Orah’. Frontiers in Plant Science, 14, 1098042.

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Prussian Blue Staining of Leishmania

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For staining with Prussian blue (Sigma-Aldrich, Germany), promastigote and intracellular amastigotes were treated with 100 µg/mL of SPIONs for 24 h. The promastigotes (control and treated cells) were washed in phosphate-buffered saline (PBS) pH 7.2 and adhered for 10 min on glass coverslips previously coated with poly-ʟ-lysine (Sigma-Aldrich, Germany). The intracellular amastigotes were obtained after infection of RAW 264.7 macrophages at a ratio of ten parasites to one macrophage. After treatment, cells were washed in PBS pH 7.2, fixed, and dehydrated, as described in [9 (link)]. Finally, cells were observed using a DM2500 optical microscope (Leica Microsystem, Germany) in bright-field mode.

Verçoza B.R., Bernardo R.R., de Oliveira L.A, & Rodrigues J.C. (2023). Green SPIONs as a novel highly selective treatment for leishmaniasis: an in vitro study against Leishmania amazonensis intracellular amastigotes. Beilstein Journal of Nanotechnology, 14, 893-903.

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SSEA-1 Immunofluorescence and Flow Cytometry

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Cells were washed twice and suspended in ice-cold PBS lacking Ca²⁺/Mg²⁺ (Gibco^®). The cell suspension (5 × 10⁴ cells) was placed onto a Superfrost™ Plus slide (Thermo Scientific™, USA). After incubation for 20 min at an ambient temperature, cells had adhered to the slide and were examined by microscopy. The remaining cell suspension was analyzed by flow cytometry. For immunofluorescence staining of stage-specific embryonic antigen-1 (SSEA-1), cells attached to a slide or in suspension (5 × 10⁵ cells) were incubated overnight at 4°C with 0.125 μg of an anti-SSEA-1 Alexa Fluor^® 488-conjugated antibody or a mouse IgM isotype control FITC-conjugated antibody (eBioscience, USA) in 500 μL of blocking buffer (Dulbecco’s PBS containing 1% bovine serum albumin (Sigma-Aldrich)). Slides were washed with PBS and mounted using ProLong™ Gold Antifade Mountant containing DAPI (Life Technologies, USA). Images were acquired using a Leica DM2500 Optical Microscope (Leica Microsystems, Germany) equipped with a Canon EOS 7D camera (Canon, Japan). Flow cytometric analysis was conducted on a Cytomics FC500 cytometer (Beckman Coulter, USA). Data were analyzed using CXP software (Beckman Coulter).

Chen Y.C., Chang W.C., Lin S.P., Minami M., Jean C., Hayashi H., Rival-Gervier S., Kanaki T., Wu S.C, & Pain B. (2018). Three-dimensional culture of chicken primordial germ cells (cPGCs) in defined media containing the functional polymer FP003. PLoS ONE, 13(9), e0200515.

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Immunofluorescence Analysis of Embryonic Tissue

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E10.5 embryos were fixed in 4% (w/v) paraformaldehyde (PFA) in PBS at 4°C. Embryos were then immersed in 15% and 30% sucrose and embedded in Tissue Freezing Medium (Triangle Biomedical Sciences, Inc.). Sections were taken at 8 μm. For immunostaining sections were blocked in 10% donkey serum/1% BSA and permeabilized in 0.1% Tween-20. Primary antibodies are listed in Supplementary Table 1. Secondary antibodies were: donkey anti-mouse Alexa Fluor 488, donkey anti-mouse Alexa Fluor 594, donkey anti-rabbit Alexa Fluor 488, donkey anti-rabbit Alexa Fluor 594 (Molecular Probes), goat anti-chicken Alexa Fluor 488 (Molecular Probes), and donkey anti-rabbit Alexa Fluor 647 (Molecular Probes). Samples were mounted in ProlongGold (Molecular Probes) and images captured using a Leica DM2500 optical microscope or Leica TSC 5SP X confocal microscope (Leica Microsystems). EdU staining on paraffin embedded kidney sections was conducted using Click-iT™ EdU imaging kit (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. Animals were injected EdU at 25 mg/kg and euthanized 2 hours later for analysis. A least 3 individual kidneys were analyzed per genotype per age group. From each kidney 10 images from the cortex were taken at 200x magnification, and the total number of EdU+ nuclei and DAPI+ nuclei in the DBA+ collecting ducts were counted for calculating the EdU ratio.

Airik R., Airik M., Schueler M., Bates C.M, & Hildebrandt F. (2019). Roscovitine blocks collecting duct cyst growth in Cep164-deficient kidneys. Kidney international, 96(2), 320-326.

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Immunofluorescence Staining of hAMSCs

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To prepare the cells for immunofluorescence staining, hAMSCs were washed with 1x DPBS (Sigma, D5773) and fixed with 4% paraformaldehyde (EMPROVE®, Germany, 1,040,051,000) for 5 min at 4 °C. The cells were then washed again with 1x DPBS and permeabilized with 0.1% Triton-X100 in 1x DPBS for 5 min at 4 °C. The following primary antibodies were used for staining: anti-CD44 (1:50), anti-CD73 (1:200), anti-CD90 (1:100), anti-CD105 (1:100), anti-CD34 (1:250), anti-CD11b (1:250), anti-CD19 (1:250), anti-CD45 (1:250), and anti-HLA-DR (1:250) (Invitrogen™). Differentiating cells were stained with Sarcomeric alpha actinin monoclonal antibody (EA-53) (Invitrogen, MAI-22863, 1:500) and Anti-Cardiac Troponin T (abcam, ab8295, 1:1000) primary antibodies, Goat Anti-Mouse IgG H&L secondary antibody (abcam, ab150113, 1:1000), and DAPI (Biotium, 40043, 1:1000) to visualize nuclei. The antibodies were diluted in PBS containing 1% bovine serum albumin (Sigma, Germany, A2153) and cells were incubated with the primary antibodies overnight at 4 °C. The cells were then washed with PBS. To observe the results of immunofluorescence staining in cardiac genes, we used Leica DM2500 Optical Microscope (Leica Microsystems, Germany) equipped with a Canon EOS 7D camera (Canon, Japan). As part of the cardiac gene expression analysis, human cardiomyocytes (Promocell, C-12810) were used as positive control.

Shih H.M., Chen Y.C., Yeh Y.T., Peng F.S, & Wu S.C. (2024). Assessment of the feasibility of human amniotic membrane stem cell-derived cardiomyocytes in vitro. Heliyon, 10(7), e28398.

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Identification and Isolation of Earthworm Astome Ciliates

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Lumbricid earthworms were collected at 25 localities in western Slovakia (Central Europe) especially in the capital city and its vicinity (Supplementary Table 1). Earthworms were determined based on the features of their external morphology (Pižl, 2002 ). The morphological identification was verified using NADH-ubiquinone oxidoreductase chain 1 (ND1) sequences, which were blasted against the National Center for Biotechnology Information (NCBI) database. The determination of earthworms from which astome ciliates had been isolated was further confirmed using the barcoding cytochrome c oxidase subunit I (COI). Primers and PCR conditions used for the amplification of ND1 and COI genes of earthworms are provided in Supplementary Tables 2, 3. The molecular assignment of examined earthworms to species is shown in Supplementary Figures 1, 2.
Earthworms were processed and dissected as described by Obert and Vd’ačný (2019 (link), 2020a) (link). Detected astome ciliates were manually isolated from the gut content of their earthworm hosts with Pasteur micropipettes adjusted as described by Foissner (2014) (link). Living ciliates were investigated in vivo at low (50–400 ×) and high (1000 ×, oil immersion) magnifications with bright field and differential interference contrast under a Leica DM2500 optical microscope (Leica Microsystems GmbH, Wetzlar, Germany).

Obert T., Rurik I, & Vd’ačný P. (2021). Diversity and Eco-Evolutionary Associations of Endosymbiotic Astome Ciliates With Their Lumbricid Earthworm Hosts. Frontiers in Microbiology, 12, 689987.

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Immunofluorescent Staining of Colon Tissue

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Colon tissue samples from three groups (vehicle, TCDD, and TCDD+Sch527123) were fixed in 4% paraformaldehyde diluted in PBS overnight. Fixed colons were sectioned (5 µm thick) and placed on coated slides. Slides were incubated for 30 min in glycine 0.1% and 1× Triton for tissue permeabilization for Arg1 staining or with glycine 0.1% only for CD11b and Gr-1 staining. Slides were then incubated with primary mouse anti-Arg1 antibodies or primary rat anti-CD11b and anti-Gr-1 antibodies purchased from Cell Signaling (Danvers, MA, USA) at 4 °C overnight, followed by a 2 h incubation at room temperature with secondary Alexa Fluor 488 goat anti-mouse IgG antibody for Arg1, Alexa Fluor 488 goat anti-rat IgG for CD11b, and Cy5 goat anti-rat IgG antibody for Gr-1. Fluorescent imaging of colon sections was taken using a Leica DM 2500 optical microscope from Leica Microsystems (Buffalo Grove, IL, USA). The quantification of cell markers was calculated as corrected total cell fluorescence (CTCF) using Image J software (National Institutes of Health and the Laboratory for Optical and Computational Instrumentation).

Neamah W.H., Busbee P.B., Alghetaa H., Abdulla O.A., Nagarkatti M, & Nagarkatti P. (2020). AhR Activation Leads to Alterations in the Gut Microbiome with Consequent Effect on Induction of Myeloid Derived Suppressor Cells in a CXCR2-Dependent Manner. International Journal of Molecular Sciences, 21(24), 9613.

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Trilineage Differentiation of hAMSCs

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To induce trilineage differentiation, hAMSCs (cell passage = 4) were cultured using the StemPro™ Adipogenesis Differentiation Kit, StemPro™ Chondrogenesis Differentiation Kit, and StemPro™ Osteogenesis Differentiation Kit (Gibco™, USA, A1007001, A1007101, A1007201). The cells were induced and cultured for 14, 21, and 28 days according to the manufacturer's instructions. After the differentiation period, the cells were stained with Oil Red O for adipogenic differentiation, Alcian Blue for chondrogenic differentiation, and Alizarin Red for osteogenic differentiation. We observed the results with the Leica DM2500 Optical Microscope (Leica Microsystems, Germany) equipped with a Canon EOS 7D camera (Canon, Japan).

Shih H.M., Chen Y.C., Yeh Y.T., Peng F.S, & Wu S.C. (2024). Assessment of the feasibility of human amniotic membrane stem cell-derived cardiomyocytes in vitro. Heliyon, 10(7), e28398.

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Placental Immunohistochemistry Assay

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Paraffin-embedded placental tissue sections from NP and preeclampsia placentas were cut at 8 μm and attached onto slides. Tissues were stored at room temperature until use. Sections were heated at 62°C for 30 minutes, immersed in Histo-Clear (National Diagnostics, Atlanta) and rehydrated using ethanol. Antigens were retrieved using 10 mmol/L sodium citrate buffer, and endogenous peroxidase activity was blocked with 3% v/v hydrogen peroxide. The sections were treated with blocking agent 10% v/v fetal calf serum in PBS for 1 hour and incubated overnight at 4°C with mouse monoclonal antibodies: anti-NEP (1:25, ab951; Abcam, Cambridge, United Kingdom) and anti-IgG1 (1:25, 400153; BioLegend, CA). VECTASTAIN Elite ABC Kit (Vector Laboratories, Peterborough, United Kingdom) reagents were used to incubate sections at room temperature in biotinylated horse anti-mouse IgG secondary antibody for 1 hour and enhancer reagent for 30 minutes. Exposure to DAB Substrate Kit (Vector Laboratories) was used to detect specific antibody binding, and nuclei were stained with Shandon Harris Hematoxylin (Thermo Fisher Scientific, MA). Sections were dehydrated using ethanol, mounted with Depex (VWR, Philadelphia), and imaged using a Leica DM2500 optical microscope (Leica Microsystems, Wetzlar, Germany) with a Micropublisher 5.0 RTV camera system (QImaging, Surrey, Canada).

Gill M., Motta-Mejia C., Kandzija N., Cooke W., Zhang W., Cerdeira A.S., Bastie C., Redman C, & Vatish M. (2019). Placental Syncytiotrophoblast-Derived Extracellular Vesicles Carry Active NEP (Neprilysin) and Are Increased in Preeclampsia. Hypertension (Dallas, Tex. : 1979), 73(5).

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Multimodal Imaging Techniques for Cellular Analysis

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Images for immunocytochemistry and immunohistochemistry were acquired using a Nikon A1R confocal microscope with a NIS-Elements multiplatform acquisition software or a Zeiss 880 Airyscan confocal microscope with a Zen Black software at the Fluorescence Microscopy Core Facility of the University of Utah. Images for SEM and TEM were taken by the FEI Quanta 600 field emission gun at the University of Utah Nanofab and FEI Tecnai 12 transmission electron microscope at the University of Utah Electron Microscopy Core Laboratory, respectively. All images were processed and analyzed with the Fiji open source software (https://fiji.sc/). Images for semithin sections and in situ hybridization were obtained by a Leica DM2500 optical microscope with Leica Las software V3.8.

Kim D.K., Kim J.A., Park J., Niazi A., Almishaal A, & Park S. (2019). The release of surface-anchored α-tectorin, an apical extracellular matrix protein, mediates tectorial membrane organization. Science Advances, 5(11), eaay6300.

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