Dmr 3000

Manufactured by Leica

Sourced in Germany

The Leica DMR 3000 is a compound microscope designed for laboratory use. It features a sturdy, ergonomic design and high-quality optics to provide clear, detailed images of specimens. The DMR 3000 is capable of various magnification levels to accommodate a range of sample types and research applications.

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Lab products found in correlation

Facscalibur, by BD (3 mentions) Rabbit anti cd68 antibody, by Proteintech (1 mentions) Anti cd206 antibody, by Proteintech (1 mentions) Anti inos antibody, by Proteintech (1 mentions) 3 3 diaminobenzidine (dab), by Merck Group (1 mentions) Dapi 4 6 diamidino 2 phenylindole, by Merck Group (1 mentions) Anti cd11b antibody, by ABclonal (1 mentions) Rm2235, by Leica (1 mentions) Masson s trichrome staining, by Wuhan Servicebio Technology (1 mentions) Hematoxylin and eosin h e, by Wuhan Servicebio Technology (1 mentions) Alexa fluor 488 or 594 conjugated secondary antibodies, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) H e stain kit, by Beyotime (1 mentions) Masson stain kit, by Solarbio (1 mentions) Citrate buffer, by Agilent Technologies (1 mentions) 4 6 diamino 2 phenyl indole dapi solution, by Beyotime (1 mentions) 6 well plate, by Corning (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions) Matrigel, by BD (1 mentions)

18 protocols using dmr 3000

Immunohistochemical and Immunofluorescence Analysis of Rat Heart

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Rat hearts were excised and fixed with 4% paraformaldehyde for 24 h. For immunohistochemistry, the heart was embedded in paraffin, and 4-µm sections were prepared. The sections were incubated with the retrieval repair solution in a water bath for 30 min after deparaffinization, followed by blocking of endogenous peroxidase with 3% H₂O₂ for 15 min. After incubation with goat serum for 2 h, sections were treated with a rabbit anti-CD68 antibody (1:200, Proteintech, Chicago, IL, USA) at 4 °C overnight. Finally, the sections were stained with 3,3'-diaminobenzidine (DAB; Sigma-Aldrich).
For immunofluorescence analysis, the heart was dehydrated with gradient sucrose (10%, 20% and 30%). The samples were frozen and cut into 12-µm slices and dried for 24 h. Samples were incubated with primary antibody overnight at 4 °C, such as anti-Ccl2 antibody (1:200, EMD Millipore, Temecula, CA, USA), anti-CD206 antibody (1:200, Proteintech), anti-iNOS antibody (1:200, Proteintech), anti-CD14 antibody (1:200, Proteintech), and anti-CD11b antibody (1:200, ABclonal Technology Co.,Ltd, Wuhan, China). After being washed 3 times with PBS, the slices were stained with DAPI (4',6-diamidino-2-phenylindole, Sigma-Aldrich) and analyzed with a fluorescence microscope (Leica DMR 3000; Leica, Bensheim, Germany) to detect fluorescein isothiocyanate (FITC)-positive signals.

Huang Z., Li X., Zhou T., Jiang Y, & Shi J.H. (2021). Phosphorylated nuclear factor erythroid 2-related factor 2 promotes the secretion of C-C motif chemokine ligand 2 and the recruitment of M2 macrophages. Annals of Translational Medicine, 9(23), 1719.

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Immunohistochemical Analysis of PRKCB in LUAD

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IHC method was used to detect the expression level of PRKCB in paraffin‐embedded LUAD specimens. The samples were first incubated with rabbit anti‐PRKCB antibody (1:200), and then incubated with goat anti‐rabbit secondary antibody (1:500) for secondary staining. Finally, a microscope (Leica DMR 3000; Leica Microsystem) was used to capture images of each slice at a magnification of 200‐fold. PRKCB (brown)‐positive staining is mainly located in the cytoplasm. It was scored according to staining intensity and percentage of PRKCB‐positive tumor cells. The median was used as the cutoff value for high or low PRKCB expression.

Wang J., Shi M., Zhang H., Zhou H., Huang Z., Zhou Y, & Shi J. (2022). PRKCB is relevant to prognosis of lung adenocarcinoma through methylation and immune infiltration. Thoracic Cancer, 13(12), 1837-1849.

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Histological and Immunohistochemical Analysis of Tissue Samples

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The acquired sample segments were submerged in 10% paraformaldehyde for 24 h. A paraffin microtome (RM2235, Leica, Wetazlar, GER) was used to slice samples embedded in paraffin blocks into 5-m thick sections.The slides were rehydrated dried and given a PBS washing. The slides were subsequently stained with hematoxylin and eosin (HE) (Servicebio, Wuhan, China) for histological analysis to gauge the percentage of inflammatory cells. Masson’s trichrome staining (Servicebio) was used to examine the density of collagen fibers and variations in collagen under a microscope.Subsequently, the slides were then treated with rabbit anti-ASPN antibody, rabbit anti-SMA antibody, and rabbit anti-collagen I antibody overnight at 4°C each and incubated with the secondary antibody at room temperature for 2 h. The slides were next stained with Meyer’s hematoxylin and 3,3-diaminobenzidine (DAB; Sigma, St. Louis, Missouri, United States) for 1–2 min (Sorabio, Beijing, P.R.C.). After blocking with neutral gum (Invitrogen, San Diego, CA, United States), the slides were examined under a microscope (Leica DMR 3000; Leica, Bensheim, Germany). ImageJ was used to quantitatively analyse the immunohistochemistry image data.

Dong Y., Zhang C., Zhang Q., Li Z., Wang Y., Yan J., Wu G., Qiu L., Zhu Z., Wang B., Gu H, & Zhang Y. (2022). Identification of nanoparticle-mediated siRNA-ASPN as a key gene target in the treatment of keloids. Frontiers in Bioengineering and Biotechnology, 10, 1025546.

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Histological Assessment of Skin Healing

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At the end of the experiment, the rats were anesthetized by sodium thiopental (40 mg/kg IP) after 12 h of fasting. Then, vascular perfusion fixation through the left ventricle was performed. The rats were fixed on an operating table to take skin specimens. The half of the skin tissue collected from rats in all groups was fixed in 10% buffered formol saline, embedded in paraffin, and sectioned at 4.0 μm. The sections were dehydrated with successive concentrations of ethanol and washed twice in distilled water. The sections of the skin tissue at days 7 and 14 were stained with hematoxylin and eosin (H&E) and with Masson’s trichrome in accordance with the protocols of the manufacturer to detect the reepithelialization/granulation tissue formation and collagen deposition, respectively. Finally, the histological sections were observed and analyzed under a microscope (Leica DMR 3000; Leica Microsystem) by two blinded experienced investigators [34 ].

Ebrahim N., Dessouky A.A., Mostafa O., Hassouna A., Yousef M.M., Seleem Y., El Gebaly E.A., Allam M.M., Farid A.S., Saffaf B.A., Sabry D., Nawar A., Shoulah A.A., Khalil A.H., Abdalla S.F., El-Sherbiny M., Elsherbiny N.M, & Salim R.F. (2021). Adipose mesenchymal stem cells combined with platelet-rich plasma accelerate diabetic wound healing by modulating the Notch pathway. Stem Cell Research & Therapy, 12, 392.

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Histological Analysis of Cardiac Tissue

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Cardiac tissues were excised and fixed in buffered formol saline (10%). Fixed tissue specimens were used for the preparation of 4–6 µm thick paraffin sections, which were mounted on glass slides for hematoxylin and eosin (H&E) and Masson’s trichrome stains, and positively charged slides were prepared for immunohistochemical (IHC) staining.
For H&E and Masson’s trichrome staining, fixed sections were dehydrated with successive concentrations of ethanol and washed twice in distilled water, followed by staining with H&E and Masson’s trichrome stains. Then, the histological sections were observed, analyzed, and imaged by two blinded experienced investigators using a light microscope (Leica DMR 3000; Leica Microsystem, Wetzlar, Germany) [95 (link)].

Ebrahim N., Al Saihati H.A., Mostafa O., Hassouna A., Abdulsamea S., Abd El Aziz M. El Gebaly E., Abo-Rayah N.H., Sabry D., El-Sherbiny M., Madboly A.G., Hussien N.I., Saadani R.E., Ebrahim H.A., Badr O.A., Elsherbiny N.M, & Salim R.F. (2022). Prophylactic Evidence of MSCs-Derived Exosomes in Doxorubicin/Trastuzumab-Induced Cardiotoxicity: Beyond Mechanistic Target of NRG-1/Erb Signaling Pathway. International Journal of Molecular Sciences, 23(11), 5967.

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Immunofluorescent Visualization of Nrf2 and PRDX5

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Immunofluorescence assay according the following steps: first the cells fixed in 4% paraformaldehyde for 0.5 hour at room temperature, PBS buffer washing for three times, and blocked with 5% donkey serum for 2 hours at room temperature, then incubated with anti-Nrf2 and anti-PRDX5 antibodies (Abcam) at 4°C overnight. PBS buffer washing for three times, the cells then were incubated with Alexa Fluor 488 or 594 conjugated secondary antibodies (Abcam) for 1 hour at room temperature. DAPI (Sigma-Aldrich) was used for nuclear staining. After adding anti-fluorescence quenching agent seal, the cells were visualized by the fluorescence microscopy (Leica DMR 3000; Leica Microsystem, Bensheim, Germany).

Chen X., Cao X., Xiao W., Li B, & Xue Q. (2020). PRDX5 as a novel binding partner in Nrf2-mediated NSCLC progression under oxidative stress. Aging (Albany NY), 12(1), 122-137.

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Clonogenic Assay Protocol

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At 24 h post-transfection, cells were seeded into 6-well plates and cultured for two weeks in DMEM containing 12% FBS at 37°C in a humiliated atmosphere of 5% CO₂ (600 cells per well in 6-well plates). The colonies were fixed and stained with 1 mg/ml crystal violet, and colonies containing >50 cells were counted using Leica microscopy (Leica DMR 3000; Leica Microsystems GmbH).

Dong Y., Zhang N., Zhao S., Chen X., Li F, & Tao X. (2019). miR-221-3p and miR-15b-5p promote cell proliferation and invasion by targeting Axin2 in liver cancer. Oncology Letters, 18(6), 6491-6500.

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Histological Analysis of Skin Appendages

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Eight rats were sacrificed randomly in each group in the first 4 weeks after injury, and the wound tissue was collected and placed in 10% formalin solution. The samples were dehydrated, embedded in 10% formalin solution overnight, sliced into sections (4 µm) and stained with H&E. A microscope (Leica DMR 3000; Leica Microsystems, Bensheim, Germany) was used to observe the changes in the skin appendages (sebaceous glands and hair follicles) and the growth of new epithelium.

Zhang C., Zhang Q., Yang D., Qiao Y., Wang B., Yan J., Li Z., Huang Z., Zhou Y., Hu K, & Zhang Y. (2022). Chitosan degradation products promote healing of burn wounds of rat skin. Frontiers in Bioengineering and Biotechnology, 10, 1002437.

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ASPN siRNA Transfection Efficiency in Keloid Fibroblasts

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To investigate the transfection effectiveness of the of the ASPN siRNA/nanoparticle (si-ASPN) complex in keloid fibroblasts in vitro. Cells were seeded in 24-well plates until roughly 70%–80% coverage was reached in each well. Using a fluorescent microscope (Leica DMR 3000; Leica Microsystem, Bensheim, GER) equipped with GFP channels, fibroblasts were grown in non-transfected (Control) or ASPN siRNA/nanoparticle (si-ASPN) mixture for 24 h (excitation 488 nm, emission 507 nm). Then, the cells were collected in plates, suspended in PBS, and a FACSCalibur flow cytometer was used to assess the transfection efficiency (BD FACSCalibur, BD Bioscience, San Jose, CA, United States).

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Histological Examination of Mouse Lungs

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The left lung lobes of mice were fixed with 4% paraformaldehyde for 24 hours. Then, the lungs exposed to gradient alcohol were embedded in paraffin and cut into 4 mm sections onto polylysine-coated slides. The sections were then deparaffinized in xylene and rehydrated with graded alcohol. Next, we performed staining using a Masson stain kit (Solarbio, Beijing, China) and a HE stain kit (Beyotime, Jiangsu, China) according to the manufacturer’s instructions. The lung sections were then observed under a microscope (Leica DMR 3000; Leica Microsystem, Bensheim, Germany).

Wang S., Huang Z., Liu Y., Sun H., Zhou Y, & Shi J. (2023). Sustainably released nanoparticle-based rhynchophylline limits pulmonary fibrosis by inhibiting the TEK-PI3K/AKT signaling pathway. Translational Lung Cancer Research, 12(3), 427-445.

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