For the wound-healing assay, confluent prostate cell monolayers were wounded by scratching a pipette tip from the top to the bottom of the well. Then images were taken of the whole length of the wound at the indicated time points using a Leica DMI 3000 B microscope. For cell invasion assay, cells in media containing 2% FBS were seeded into the top chamber of an 8 µm pore transwell insert (Corning, NY) coated with Matrigel (BD Biosciences) (1 × 104 cells/well/100 µl) in a 24-well plate. Medium containing 20% FBS was added to the bottom chamber and cells were allowed to migrate for indicated time. Cells that had not migrated were wiped from the top of the chamber. The chambers were fixed with 10% TCA, stained with 0.4% SRB (Sulforhodamine B) (Sigma, USA) for 30 min and washed five times with 1% acetic acid. Images were taken using a Leica DMI 3000 B microscope.
Dmi3000b microscope
Manufactured by Leica
Sourced in Germany, United Kingdom, United States
The DMI3000B is a high-quality inverted microscope designed for versatile laboratory applications. It features a robust and stable construction, as well as a modular design that allows for a wide range of configurations and accessories. The DMI3000B microscope provides reliable performance and is suitable for various scientific and research purposes.
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Lab products found in correlation
Tissue tek tec 5, by Sakura Finetek (5 mentions)
Dfc3000g camera, by Leica (4 mentions)
Matrigel, by BD (4 mentions)
Nova lite hep 2 ana kit, by Inova Diagnostics (4 mentions)
Edu imaging kit, by Thermo Fisher Scientific (4 mentions)
Microplate reader, by Thermo Fisher Scientific (4 mentions)
Tissue tek vip 6, by Sakura Finetek (3 mentions)
Fv1000 confocal microscope, by Olympus (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
Sirius red stain kit, by Beyotime (3 mentions)
Alexa fluor 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (3 mentions)
Cck 8, by Beyotime (3 mentions)
Micromanipulator, by Leica (3 mentions)
Envision multilabel reader, by PerkinElmer (2 mentions)
Metamorph software, by Molecular Devices (2 mentions)
Dfc450 camera, by Leica (2 mentions)
Fluoroshield mounting medium with dapi, by Abcam (2 mentions)
Las v 4, by Leica (2 mentions)
Sulforhodamine b srb, by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (2 mentions)
170 protocols using dmi3000b microscope
1
Wound Healing and Cell Invasion Assay
2
Tube Formation and Wound Healing Assays
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For tube formation experiments, human umbilical vein endothelial cells (HUVECs) were digested with 0.25% trypsin and seeded in Matrigel-coated 48-well plates at 1 × 105 cells per well with 200 μL conditioned medium from modLuciferase-transfected or modVEGFA-transfected fibroblasts on CEMC. Tube formation was visualized and photographed every 2 h after treatment using a DMI3000B microscope (Leica). The number of tubes and junctions was counted using Image-pro plus 6.0 (Media Cybernetics). All experiments were performed independently with five replicates per group.
For the wound healing assays, 5 × 105 HUVECs were cultured in each well of a 6-well plate. Straight scratches were made in confluent HUVEC monolayers with a sterile 100 μL pipette tip and then incubated with conditioned medium collected from modLuciferase-transfected or modVEGFA-transfected fibroblasts on CEMC for 48 h. Images were captured with a DMI3000B microscope (Leica) at 0 h, 24 h, and 48 h and analyzed using Image-pro plus 6.0 (Media Cybernetics). All experiments were conducted with five replicates per group.
For the wound healing assays, 5 × 105 HUVECs were cultured in each well of a 6-well plate. Straight scratches were made in confluent HUVEC monolayers with a sterile 100 μL pipette tip and then incubated with conditioned medium collected from modLuciferase-transfected or modVEGFA-transfected fibroblasts on CEMC for 48 h. Images were captured with a DMI3000B microscope (Leica) at 0 h, 24 h, and 48 h and analyzed using Image-pro plus 6.0 (Media Cybernetics). All experiments were conducted with five replicates per group.
3
Capsule Induction and Quantification Protocol
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Capsule induction was performed following the protocol described by Zaragoza et al. using capsule-inducing medium (10% SDB in 50 mM MOPS, pH 7.3) [34 (link)]. An inoculum was adjusted to 1 × 108 cells/mL prior to plating. The cultures were incubated overnight at 37 °C with shaking (110 rpm). Following microscopy observation, a microscope slide containing one drop of India ink and one drop of each isolate was prepared and observed using a Leica DMI 3000B microscope (Leica Microsystems, Wetzlar, Germany). Images were acquired in bright field with 40× or 63× objectives, and capsule size and cell body (delimited by cell wall) were measured in 50 cells using ImageJ Software (Fiji) V2.1 [35 (link)]. The capsule area of 50 cells was measured for each isolate, and the procedure previously described was used to measure capsule size.
4
Wound Healing Assay with Baicalin Treatment
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Wound-healing assays were conducted as previously described with some modifications [14 (link)]. Briefly, MESO924 cells were trypsinized, seeded in 6-well plates, and cultured with 1640 basic medium supplemented with 10% FBS. Using the tip of a 200 μL pipette, a scratch was produced in the cultures that were close to fusion, then cultured for 48 h after baicalin treatment with different concentrations (50 and 100 μM). Cells were imaged at 48 h with a Leica DMI 3000B microscope (Leica Microsystems, Germany).
5
Histological Analysis of Enucleated Eyes
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Before enucleation, the superior edge of the eye was marked. Once enucleated, eyes were placed immediately in PREFER, a glyoxal fixative (Anatech Ltd, Battle Creek, MI, USA) and incubated overnight at room temperature. Eyes were then placed in cassettes and stored in 70% ethanol at room temperature. Orientated eyes were processed and embedded in paraffin for sectioning (Tissue-Tek VIP 6, Tissue-Tek TEC 5; Sakura Finetek USA, Inc., Torrance, CA, USA). Sections were cut with a microtome to a thickness of 4 μm, stained with hematoxylin-eosin (H&E), and viewed on a Leica DMI3000 B microscope (Leica Microsystems GmbH, Wetzlar, Germany). All images were taken at a magnification of ×40.
6
Oxidative Stress Evaluation in HepG2-SR Cells
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HepG2-SR cells seeded in black 96-well plates (3,000 cells/well) overnight were treated with a fluorescent dye (10 μM DCF-DA; 6-Carboxy-2′,7′-dichlorofluorescein diacetate) for 45 min. The supernatant was then discarded and replaced with fresh medium. Afterwards, the cells were treated with 75 μM Rg3, 15 μM ART or 75 μM Rg3 + 15 μM ART. Fluorescence was measured using a fluorescence microplate reader (EnVision® Multilabel Reader, PerkinElmer) at various time points at 37 °C [26 (link)]. For DCF-DA fluorescence microscopic analyses, HepG2-SR cells seeded in 6-well plate (1 × 105cells/well) were treated with 75 μM Rg3 + 15 μM ART and/or 5 mM NAC for 8 hrs. The cells were then treated with 10 μM DCF-DA for 45 min. Afterwards, samples were rinsed three times with PBS, and then counter-stained with DAPI [26 (link)]. Images were obtained by a Leica DMI3000 B microscope (Leica Microsystems, Wetzlar, Germany).
7
Photoreceptor Nuclei Quantification in Mouse Retina
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Seven days after light exposure, mouse eyes were enucleated and placed immediately in 4% paraformaldehyde for incubation overnight at room temperature. Eyes then were placed in cassettes and stored in 70% ethanol at room temperature. Oriented eyes were processed and embedded in paraffin for sectioning (Tissue-Tek VIP 6, Tissue-Tek TEC 5; Sakura Finetek USA, Inc., Torrance, CA, USA). Sections were cut with a microtome to 4 μm thick, stained with hematoxylin and eosin (H&E) and viewed on a Leica DMI3000 B microscope (Leica Microsystems GmbH, Wetzlar, Germany). Central retina (located by the optic nerve head) was imaged from three different mice in each group (PBS + No LIR, MMF + LIR, and PBS + LIR). Photoreceptor nuclei in a ×40 frame were counted and reported.
8
Wound Healing Assay with Inhibitors
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The cells were grown in silicone Culture-Inserts 4 Well (80469, Ibidi, Gräfelfing, Germany) to create the cell-free area (wound). After reaching confluence, the inserts were removed and cells washed with PBS before preincubation with the inhibitors for 30 min and stimulated with LPA or EtOH. Pictures were taken at a 10x magnification from four different fixed areas with a Leica DMI3000B microscope (Leica Microsystems, Wetzlar, Germany) using Leica Application Suite (LAS) V4.7 at time points 0 and 8 h. The cell-free area was analyzed and calculated using ImageJ (NIH) software.
9
Measuring Mitochondrial Membrane Potential in ESC-CMs
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The ΔΨm of ESC-CMs was measured as previously described [26 (link)] following the manufacturer’s instructions. ESC-CMs on d 5 + 3 were plated on 48-well plates at a density of 3 × 104 cells per well. After 48 h, the cells were incubated with 2 μg·mL−1 JC-1 dye (Beyotime) at 37 °C in the dark for 30 min. Then, the cells were washed with washing buffer, and images were obtained with a Leica DMI3000B microscope (Leica Microsystems, Wetzlar, Germany).
10
Biofilm-Induced Keratinocyte Analysis
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After 2 and 24 h of contact with soluble factors from biofilm, keratinocytes were analysed and photographed by brightfield microscopy using a Leica DMI 3000B microscope (Leica Microsystems, Wetzlar, Germany) soluble factors from biofilm.
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