AG-1S electronic universal mechanical testing machine (Shimadzu), PI3K, p-AKT, Bax, caspase-3 and tumor necrosis factor-α (TNF-α) primary antibodies (Abcam), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay kit (Shanghai Beyotime Biotechnology), quantitative polymerase chain reaction (qPCR) kit (Vazyme), immunohistochemistry kit (Maxim), bicinchoninic acid (BCA) protein quantification kit (Shanghai Beyotime Biotechnology), fluorescence qPCR instrument (ABI 7500; Applied Biosystems; Thermo Fisher Scientific, Inc.), Image-Lab image analysis system (Bio-Rad Laboratories), and Leica DM4000B LED microscope (Leica Microsystems GmbH).
Dm4000 b led microscope
Manufactured by Leica
Sourced in Germany, Belgium
The Leica DM4000 B LED microscope is a high-performance optical microscope designed for a wide range of applications. It features a LED illumination system, providing consistent and long-lasting illumination. The microscope offers precise and accurate focusing, allowing users to examine samples with exceptional clarity and detail.
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Lab products found in correlation
Las v 4, by Leica (3 mentions)
Dfc7000 t digital camera, by Leica (2 mentions)
Toluidine blue, by BioVitrum (2 mentions)
Bond max, by Leica (2 mentions)
Bond polymer refine detection kit, by Leica (2 mentions)
M0821, by Agilent Technologies (2 mentions)
Dmc2900, by Leica (2 mentions)
Eclipse ni e microscope, by Nikon (2 mentions)
Dfc450c, by Leica (2 mentions)
Tunel assay kit, by Beyotime (1 mentions)
Bicinchoninic acid bca protein quantification kit, by Beyotime (1 mentions)
Abi 7500, by Thermo Fisher Scientific (1 mentions)
P akt, by Abcam (1 mentions)
Image lab image analysis system, by Bio-Rad (1 mentions)
Dfc310 fx, by Leica (1 mentions)
Prolong gold with dapi, by Thermo Fisher Scientific (1 mentions)
Alexa 594, by Thermo Fisher Scientific (1 mentions)
Anti e cadherin, by Abcam (1 mentions)
Matlab, by MathWorks (1 mentions)
Originpro 2021b, by OriginLab (1 mentions)
38 protocols using dm4000 b led microscope
1
Evaluation of Apoptotic Signaling Pathways
2
Quantification of Myocardial Fibrosis
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After RHC, animals were sacrificed. RV and LV plus septum were then dissected for tissue weight measurement. Biopsies of RV free wall tissues were fixed in 10% buffered formalin. The fixed tissues were then embedded in paraffin and stained with picrosirius red, for measurement of collagen deposition.
Images of RV stained with picrosirius red were taken by a scientist who was blinded to the experimental groups, using a Leica digital color camera (Leica DFC310 FX, Leica Microsystems; Wetzlar, Germany) and Leica DM4000 B LED microscope with a 20X objective (Leica Microsystems; Wetzlar, Germany). For each sample, more than 4 areas were imaged and analyzed for the percentage of the collagen area using Leica software (LAS V4.7, Leica Microsystems; Wetzlar, Germany). Results are presented as the average percentage of all the sample areas stained with picrosirius red.
Images of RV stained with picrosirius red were taken by a scientist who was blinded to the experimental groups, using a Leica digital color camera (Leica DFC310 FX, Leica Microsystems; Wetzlar, Germany) and Leica DM4000 B LED microscope with a 20X objective (Leica Microsystems; Wetzlar, Germany). For each sample, more than 4 areas were imaged and analyzed for the percentage of the collagen area using Leica software (LAS V4.7, Leica Microsystems; Wetzlar, Germany). Results are presented as the average percentage of all the sample areas stained with picrosirius red.
3
Quantifying Skin Collagen Density
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Trichrome stained sections were imaged in brightfield mode, with a 20× objective, on a Leica DM4000 B LED microscope (Leica Microsystems, Wetzlar, Germany). To measure the collagen density in the skin, each section was imaged over the length of the section requiring ten evenly spaced fields of view. Using ImageJ software, the region of interest (the dermis, excluding hair follicles, sweat glands, blood vessels, and pockets of red blood cells) was selected so that only the area containing collagen was included in the analysis. Next, thresholding was used to select only blue pixels (collagen) and excluded purple/red pixels (immune cells and keratin); white hues were excluded to eliminate holes in the tissue. The collagen density was calculated as the number of pixels representing collagen divided by the total number of pixels in the region of interest (ROI). The percent area of tissue comprised of collagen was averaged for each animal and the mean per group reported.
4
Histological Analysis of Wound Healing
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The animals were sacrificed on days 4, 7, 14 and 21 after the operation, and the wound tissues were excised for histological analysis. Then, 4 μm-thick sections of the formalin-fixed, paraffin-embedded tissue samples were stained with hematoxylin and eosin (H & E) (BioVitrum, St. Petersburg, Russia), picrosirius red (BioVitrum, St. Petersburg, Russia), toluidine blue (BioVitrum, St. Petersburg, Russia) and by picro-Mallory (BioVitrum, St. Petersburg, Russia). A LEICA DM4000 B LED microscope equipped with a LEICA DFC7000 T digital camera running under the LAS V4.8 software (Leica Microsystems, Wetzlar, Germany) was used for the examination of the samples. Sections stained with picrosirius red were examined by polarized light microscopy. The panels were composed of microphotographs of central parts of the wounds for standardized assessment of wound healing.
5
Immunofluorescence Analysis of Ovarian Cancer Cells
6
Erythrocyte Transport in Microchannels
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Data for the analysis of erythrocyte transport in the microchannels were obtained by recording videos at 400–450 fps with a XIMEA MC023MG-SY camera (XIMEA GmbH, Münster, Germany) using a Leica DM4000B LED microscope (Leica Microsystems GmbH, Wetzlar, Germany) with 20×/0.40 objective lens (Leica Microsystems, Wetzlar, Germany). A region of interest containing one microchannel was selected to make the recording. During every experiment for each concentration of tBuOOH, 9–10 channels were recorded. The number of cells analyzed in each type of microchannels at each concentration of tBuOOH is presented in Table 1 .
A custom script developed in the MATLAB software package (The MathWorks) was used to calculate the velocity of RBCs in the microchannels. It determined the frames in which the cell entered and exited a channel. The difference in frames was used to calculate the velocity of the cell passage. This velocity was normalized to the velocity of the fluid flow in the channel measured by analyzing cells velocities in a wide channel before and the measurement microchannel. The data of each experiment were processed in OriginPro 2021b (OriginLab Corporation). First, the data were converted to the form of probability density (Statistics—Frequency Counts), and then they were averaged and approximated by the Gaussian function.
A custom script developed in the MATLAB software package (The MathWorks) was used to calculate the velocity of RBCs in the microchannels. It determined the frames in which the cell entered and exited a channel. The difference in frames was used to calculate the velocity of the cell passage. This velocity was normalized to the velocity of the fluid flow in the channel measured by analyzing cells velocities in a wide channel before and the measurement microchannel. The data of each experiment were processed in OriginPro 2021b (OriginLab Corporation). First, the data were converted to the form of probability density (Statistics—Frequency Counts), and then they were averaged and approximated by the Gaussian function.
7
Histological Tissue Analysis Protocol
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The samples were fixed in formalin for 24 h, then underwent standard histological processing and were embedded into paraffin blocks. Four μm thick sections of the tissue samples were stained with hematoxylin and eosin (H&E), Mallory’s trichrome stain and Picrosirius red (PSR) for the detection of collagen fibers. A LEICA DM4000 B LED microscope, equipped with a LEICA DFC7000 T digital camera running under the LAS V4.8 software (Leica Microsystems, Wetzlar, Germany) was used for the examination and imaging of the samples. The specimens were studied using standard (for H&E, Mallory and PSR stained samples) and polarized light (PSR stained samples) microscopies.
8
Immunofluorescence Staining of IL-13Rα1 and IL-4Rα
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Cells were fixed with 4% PFA and blocked for 1 h at RT with 10% protein block in PBS. Anti-IL-13Rα1 and anti-IL-4Rα (1:250, ab79277, Abcam, and 1:50, NBP1-00884, Novus Biologicals) antibodies were diluted in PBS with 1% protein block and were incubated overnight at 4 °C. Following washing, secondary antibody (donkey anti-rabbit IgG 555, 1:500) incubation was done for 1 h at RT. The specificity of the secondary antibody was tested by omitting the primary antibody. Counterstaining with DAPI was performed for 10 min. Pictures were taken using a LEICA DM4000 B LED microscope and LAS X software (Leica Microsystems).
9
EDU Staining for Cell Proliferation
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EdU (5-ethynyl-2′-deoxyuridine) staining enables quick and effective cell proliferation assays to accurately measure the proportion of cells in the S phase. Therefore, using EdU staining in accordance with the manufacturer’s instructions, the proliferation of pGCs was determined. After treatment with 0 mmol/L, 20 mmol/L, 40 mmol/L, and 60 mmol/L 2,5-HD for 24 h, cells were cultured in phenol red-free DMEM/F12 with 50 μM EdU (RiboBio, Guangzhou, China) staining for 2 h [23 (link),24 (link)]. Cells were then rinsed two times with 0.02 M PBS (pH = 7.2) at 5-min intervals before being immobilized for 30 min with 4% paraformaldehyde (Biosharp, Hefei, China). Following that, cells were treated for 5 min with glycine (Biosharp, Hefei, China) (2 mg/mL) and rinsed for 5 min with PBS [23 (link),24 (link)]. After 30 min of DAPI (0.5 μg/mL) (Biosharp, Hefei, China) staining [23 (link)], three rounds of PBS washing were conducted at 5-min intervals. Lastly, a Leica DM4000 BLED microscope (Leica Microsystems DM2500, GER) was used to observe the fluorescence in cells.
10
Autofluorescence Spectrum Analysis of Neocortical Samples
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Unstained paraffin sections of neocortical samples of four patients (E1, E2, E5, E6) and two controls (C1, C2) were used for autofluorescence spectrum analysis. A Leica DM4000 B LED microscope (Leica Microsystems GmbH, Wetzlar, Germany) was used with HC PLAN APO ×20/0.70 and HC PLAN APO ×40/0.75 water objectives, and an HXC PLAN APO ×100/1.40-0.70 oil immersion objective.
Images were recorded using a Nuance Multispectral Imaging System (PerkinElmer, Hopkinton, MA) with version 3.0.2 software.
Nuance microscopy registers the spectrum of light emitted by a specimen when excited in a narrow wavelength range. We applied excitation wavelengths between 420 and 460 nm, while emission was evaluated through a long pass filter, passing all fluorescence above 420 nm. The spectrum, obtained in ascending steps of 20 nm, was determined from several autofluorescent particles located in both the pial artery vascular wall and the neocortical tissue sections. Subsequently, multispectral analysis was performed by "unmixing" the generated spectral curves of the various fluorescent materials, thereby differentiating the spectrum of the particles of interest from the spectrum of other (background) structures. To assess potential influence of formaldehyde and/or paraffin on the spectrum, the analysis was repeated on fresh-frozen tissue in one patient (E1).
Images were recorded using a Nuance Multispectral Imaging System (PerkinElmer, Hopkinton, MA) with version 3.0.2 software.
Nuance microscopy registers the spectrum of light emitted by a specimen when excited in a narrow wavelength range. We applied excitation wavelengths between 420 and 460 nm, while emission was evaluated through a long pass filter, passing all fluorescence above 420 nm. The spectrum, obtained in ascending steps of 20 nm, was determined from several autofluorescent particles located in both the pial artery vascular wall and the neocortical tissue sections. Subsequently, multispectral analysis was performed by "unmixing" the generated spectral curves of the various fluorescent materials, thereby differentiating the spectrum of the particles of interest from the spectrum of other (background) structures. To assess potential influence of formaldehyde and/or paraffin on the spectrum, the analysis was repeated on fresh-frozen tissue in one patient (E1).
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