Liver sections were deparaffinized, rehydrated, and blocked with 5% bovine serum albumin (BSA) for 1 hour at room temperature. After washing, the tissue slices were incubated overnight at 4°C with primary rabbit antibodies against F4/80 (1:5000, Servicebio), CD206 (1:400, Servicebio), and iNOS (1:200, Servicebio). After washing with PBS, the slices were incubated at room temperature for 1 h with goat anti-rabbit secondary antibodies labeled with HRP (1:500, Servicebio) or Cy3 (1:300, Servicebio). Nuclei were stained with DAPI (Sigma-Aldrich). Fluorescence images were obtained with a DM6B microscope (Leica Microsystems, Milan, Italy) and analyzed by ImageJ.
Dm6 b microscope
Manufactured by Leica Microsystems
Sourced in Germany
The DM6 B microscope is a high-performance optical microscope from Leica Microsystems. It is designed for a wide range of applications in life science and industrial research. The DM6 B offers advanced imaging capabilities, including high-resolution, high-contrast, and high-speed imaging. It is equipped with a range of objective lenses and illumination options to accommodate various sample types and magnification requirements.
Automatically generated - may contain errors
Lab products found in correlation
Las x software, by Leica Microsystems (2 mentions)
F4 80, by Wuhan Servicebio Technology (1 mentions)
Cyanine3 (cy3), by Wuhan Servicebio Technology (1 mentions)
Cd206, by Wuhan Servicebio Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Click it plus opp alexa fluor 488 protein synthesis assay kit, by Thermo Fisher Scientific (1 mentions)
Linksys32, by Linkam (1 mentions)
Dmc6200, by Leica camera (1 mentions)
8 protocols using dm6 b microscope
1
Immunofluorescent Staining of Liver Macrophages
2
Quantifying Fungal Protein Synthesis
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To measure newly synthesized proteins in fungal cells, a Click-iT™ Plus OPP Alexa Fluor™ 488 Protein Synthesis Assay Kit (Thermo Scientific) was used according to the manufacturer’s instructions. Briefly, conidia induced on YMA were harvested and inoculated in liquid CM (1 × 106 conidia/mL). After 1 h of incubation on a rotary shaker, cells were washed with phosphate-buffered saline (PBS). Then, the cells were re-inoculated in liquid CM containing OPP with or without translation inhibitors (cycloheximide or hygromycin B) and incubated for 1 h on a rotary shaker. The cells were fixed using 3.7% formaldehyde solution and permeabilized with 0.5% Triton X-100 in PBS. Reaction cocktail containing Alexa Fluor® dye with the azide moiety was prepared for the click reaction and added to the cells. Samples were incubated for 30 min at room temperature in the dark and then washed with rinse buffer. The newly synthesized proteins were examined with a DM6 B microscope (Leica Microsystems) using the L5 filter set with a consistent exposure time. After the images had been captured, mean GFP fluorescence intensity in the transverse section of an individual conidium was quantified using LAS X software (Leica Microsystems). In total, 100 conidia from each sample were examined.
3
Fungal Macro- and Micro-Morphological Analysis
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To assess macro- and micro-morphological characters, each fungal isolate was grown on PDA at 25 ± 1°C for 14 days. The fungal spores were also obtained from PDA plates cultured for 14 days. Specimens were observed and photographed at 400 × magnification in a DM6 B microscope (Leica Microsystems, Wetzlar, Germany) equipped with a Leica DMC6200 camera.
4
Quantifying Phosphorylated α-Synuclein in SNpc
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The mice were anaesthetized and transcardially perfused with PBS. Coronal sections of the SNpc (10 μm) were incubated with rabbit anti-p-α-syn (Ser129) (1:500, 23706S; Cell Signaling Technology, Beverly, MA, USA) at 4 ℃ overnight. The slices were then incubated with biotinylated anti-rabbit IgG, treated with avidin–biotin peroxidase complex, and visualized using 0.05% DAB + 0.03% H2O2. Images were captured using a DM6 B microscope (Leica Microsystems, Wetzlar, Germany), and p-α-syn (Ser129) positive cells in the brain region were counted by ImageJ software (NIH, Bethesda, USA).
5
Quantifying Conidium Production and Morphology
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After incubating a cultured plug of CM in 5 mL CMC medium for 5 days at 25 °C on a rotary shaker (200 rpm), conidium production was evaluated by counting the numbers of conidia using a hemocytometer (Superior, Marienfeld, Germany). To observe conidium morphology, conidia induced on YMA were harvested and differential interference contrast (DIC) images were obtained using a DM6 B microscope (Leica Microsystems, Wetzlar, Germany).
6
Starch Granules Birefringence Analysis
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A T95 series temperature controller coupled with a temperature controlled stage and Linksys32 software (Linkam Scientific, Tadworth, UK) fitted to a Leica DM6 B microscope with CTR6 LED controller and LASX software (Leica Microsystems, Wetzlar, Germany) was used to analyse loss of birefringence of isolated starch granules under polarized light during heating. Aliquots (2 μl) of homogenous slurries of starch in water (25 mg ml−1) were mounted onto StarFrost silane coated slides (ProSciTech, Thuringowa, QLD, Australia). The coverslip was sealed with nail polish to prevent streaming of starch granules during heating. Hot stage temperature ramp rate was 4°C per minute to a maximum of 90°C. Images were taken at 15 s intervals over 17 min. Intensity profiles created in LASX software were exported to Microsoft Excel. The initial intensity was set at 100% and profile temperatures (95, 50, and 5% of initial intensity) were calculated assuming a linear loss of intensity. Only wild-type and starch from the highest TAG lines were analyzed as these were expected to reveal the greatest differences. Starch swelling power was determined using the method of Konik-Rose et al. (2001 (link)).
7
Intracellular Delivery of Cas9-GFP
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Protoplasts and conidia were incubated with Cas9 protein fused to green fluorescent protein (GFP) (Applied Biological Materials, Canada) containing the SV40 T antigen nuclear localization sequence (NLS). One hundred fifty microliters of protoplast (5 × 105) and conidium (5 × 105) solutions were incubated with 0.5 μL Cas9 protein (50 pmol/μL; Abcam) for the translocation assay. Samples were incubated at 4°C for an hour. Microscopic observation was achieved using the DM6 B microscope (Leica Microsystems, Wetzlar, Germany), which was equipped with the Leica DMC6200 camera and used the fluorescent filter L5 (Part No. 11504166).
8
Analyzing Fungal Fluorescence Response
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Microscopic observation was performed with a DM6 B microscope (Leica Microsystems) equipped with a Leica DMC6200 camera using the fluorescent filter L5 (part no. 11504166). Conidial suspensions of the PFCR1‐GFP strain were inoculated in CM at 2 × 105 conidia/ml, and mycelia were harvested 24 hr after incubation on a rotary shaker (200 rpm). The mycelia were observed under UV light 8 and 12 hr after reinoculation in CM and CM supplemented with CuSO4 (10 or 50 μM) or BCS (25 or 50 μM).
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