In order to assess the morphological features of the yeast isolates, they were cultivated in YPD broth (Merck Millipore, Darmstadt, Germany) for 2 days on a Heidolph Titramax 1000 rotary shaker (Heidolph, Schwabach, Germany) at 125 rpm. Pulcherrimin production was evaluated in YPD broth with 0.05% FeCl3. The yeast cells and pulcherrimin were observed using an OLYMPUS BX41 light microscope (Olympus, Shinjuku, Tokio, Japan) connected to a DP72 digital camera. In order to determine growth profiles under varying environmental conditions, including temperature (2–55 °C), pH (2–9), peroxide (0–50 mM), glucose (0.5–30% w/v), and ethanol concentration (1–8% w/v), YPD medium was also used. Inoculation of the culture media was carried out using standardized suspensions of the tested yeasts [~ 1°McF]. The intensity of yeast growth was measured using a DEN-1densitometer (Grant Instruments, Cambridge, UK) and expressed on the McFarland scale [°McF].
Bx41 light microscope
Manufactured by Olympus
Sourced in Japan, United States
The BX41 is a light microscope designed for various laboratory applications. It features a sturdy optical system that provides high-quality magnification and clarity. The microscope is capable of bright-field and phase contrast observation modes.
Automatically generated - may contain errors
Lab products found in correlation
Envision flex hematoxylin, by Agilent Technologies (3 mentions)
Dp12 digital camera, by Olympus (3 mentions)
Envision flex peroxidase blocking reagent, by Agilent Technologies (2 mentions)
Dako mounting medium, by Agilent Technologies (2 mentions)
Dako autostainer link 48, by Agilent Technologies (2 mentions)
Dp2 bsw, by Olympus (2 mentions)
Autostainer xl, by Leica Biosystems (2 mentions)
Dp70 camera, by Olympus (2 mentions)
Citifluor, by Agar Scientific (2 mentions)
Propylene oxide, by Merck Group (2 mentions)
Glutaraldehyde, by Merck Group (2 mentions)
Diff quik stain set, by Siemens (2 mentions)
Ypd broth, by Merck Group (1 mentions)
Titramax 1000 rotary shaker, by Heidolph (1 mentions)
Den 1densitometer, by Grant Instruments (1 mentions)
Transwell chamber, by Corning (1 mentions)
Crystal violet, by Beyotime (1 mentions)
Analysis software, by Olympus (1 mentions)
Envision flex target retrieval solution, by Agilent Technologies (1 mentions)
Ab84952, by Abcam (1 mentions)
138 protocols using bx41 light microscope
1
Comprehensive Yeast Characterization Protocol
2
Measuring Cell Migration through Matrigel
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Cell migration through Matrigel-coated filters was measured by using Transwell chambers (Costar Corporation, Tewksbury, MA, USA) with 8-μm-pore polycarbonate filters coated with Matrigel matrix. SGC-7901 cells were seeded in the upper compartment of each invasion chamber and incubated in the presence of COS for 48 h. The lower well was filled to the top (500 μl) with RPMI-1640 containing 10% fetal calf serum (Hangzhou Sijiqing Bioengineering Material Co., Ltd., Hangzhou, China) as a chemoattractant. Subsequently, non-migrating cells on the upper surface of the membrane were removed by gently scrubbing with a cotton swab, and the invading cells on the lower surface were fixed with 100% methanol and stained with crystal violet (0.1%; Beyotime Institute of Biotechnology, Nantong, China). The number of cells was counted under a light microscope (Olympus BX41; Olympus Corporation) at a magnification of ×100. Images were captured and subjected to computer-assisted image analysis using a computer coupled to the Olympus BX41 light microscope (Olympus Corporation) using AnalySis software (Olympus Corporation).
3
Immunohistochemical Detection of AMH
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Slides from TMAs (4 μm-thick) were used for immunohistochemistry (IHC) reactions, which were performed using DakoAutostainer Link48 (Dako, Glostrup, Denmark). In order to deparaffinize, rehydrate and unmask the antigens the sections were boiled in EnVision FLEX Target Retrieval Solution (pH 9, 20 min, 97 °C; Dako) using the PTLink platform (Dako, Glostrup, Denmark). Afterwards, slides were incubated for 5 min with Envision Flex Peroxidase-Blocking Reagent (Dako, Glostrup, Denmark) to block endogenous peroxidase. As primary antibodies (20 min, RT), rabbit polyclonal antibodies against AMH (1:100, ab84952, Abcam, Cambridge, UK) were used. Next, slides were incubated with EnVision FLEX/HRP (20 min, RT), and the reaction was visualized (10 min, RT) with freshly prepared 3,3′-diaminobenzidine (DAB). Additionally, slides were counterstained for 5 min with EnVision FLEX Hematoxylin (Dako, Glostrup, Denmark). Finally, slides were dehydrated in ethanol (70%, 96%, absolute) and xylene, then mounted with Dako Mounting Medium (Dako, Glostrup, Denmark). Slides were evaluated using the Olympus BX41 light microscope (Olympus, Japan). Control tissues included the human prostate.
4
Cell Viability and Clonogenic Assays
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For the cell viability assay, the cells were seeded into 96-well microtiter plates at a density of 3,000 cells/well and allowed to adhere for 12 h. Cells were then treated with different chemicals for the indicated durations and then exposed to CCK-8 (10 µl/well) for 2 h at 37°C. Absorbance was measured at 450 nm on a Tecan Sunrise microplate reader (Tecan Group AG, Männedorf, Switzerland).
For the clonogenic assay, 800 cells were plated in triplicate in 6-well plates. After overnight incubation at 37°C, different chemicals were added into the medium to reach various concentrations (0, 25, 50, 100, 300 and 1,000 nM). Medium was changed every 3–4 days while maintaining the previous concentrations. After 9–10 days of culture, cell colonies were fixed with ice-cold methanol, followed by 0.05% crystal violet staining for 15 min. Colonies containing >50 cells in each well were counted manually under an Olympus BX41 light microscope (Olympus Corp., Tokyo, Japan). A gridded plastic sheet was attached to the bottom of each well to keep track of colonies counted (26 (link)). All fields were counted.
For the clonogenic assay, 800 cells were plated in triplicate in 6-well plates. After overnight incubation at 37°C, different chemicals were added into the medium to reach various concentrations (0, 25, 50, 100, 300 and 1,000 nM). Medium was changed every 3–4 days while maintaining the previous concentrations. After 9–10 days of culture, cell colonies were fixed with ice-cold methanol, followed by 0.05% crystal violet staining for 15 min. Colonies containing >50 cells in each well were counted manually under an Olympus BX41 light microscope (Olympus Corp., Tokyo, Japan). A gridded plastic sheet was attached to the bottom of each well to keep track of colonies counted (26 (link)). All fields were counted.
5
Transwell Assay for Cell Migration and Invasion
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After 48 h transfection, the cells were suspended in serum-free DMEM (1 × 104 cells/100 μL) and inoculated into the upper layer of transwell chamber. The lower layer was filled with medium containing 10% FBS. After maintenance at 37℃ overnight, the migrated cells were fixed and stained. The number of cells in five randomly selected microscopic vision fields was then counted and the cells were imaged under an Olympus BX41 light microscope (Olympus Corporation) at a magnification of ×200. For invasion assay, the upper layer of transwell chamber was coated with Matrigel.
6
Microscopic Evaluation of PCNA Expression
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The analysis of the preparations and their photographic documentation were performed using an Olympus BX41 light microscope (Olympus Corporation) equipped with Olympus DP12-2 camera (Olympus Corporation). Images from five randomly selected microscopic fields (each field 0.785 mm2), at a magnification of 200× were morphometrically evaluated by using image analysis software (NIS-Elements Advanced Research Nikon software, Nikon Instruments, Melville, NY, USA).
The number of PCNA-IR cells was counted in each analyzed image and presented as median per visual field (0.785 mm2). Morphological characteristics of PCNA-IR nuclei, (diameter, area), intensity of immunohistochemical reaction, and thickness of the epidermis were also evaluated. Intensity of immunohistochemical reaction was measured by using 0 to 256 gray scale level, where completely black pixel had a value of 0, and completely white or bright one possessed a value of 256. The gray scale intensity and PCNA expression have inverse relation.
The number of PCNA-IR cells was counted in each analyzed image and presented as median per visual field (0.785 mm2). Morphological characteristics of PCNA-IR nuclei, (diameter, area), intensity of immunohistochemical reaction, and thickness of the epidermis were also evaluated. Intensity of immunohistochemical reaction was measured by using 0 to 256 gray scale level, where completely black pixel had a value of 0, and completely white or bright one possessed a value of 256. The gray scale intensity and PCNA expression have inverse relation.
7
Staining and Microscopic Analysis of Plant Roots
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Plant roots were harvested and stained according to a modified protocol of the ink-vinegar technique of Vierheilig et al. (1998) (link). Roots were first rinsed under running tap water to remove adhering soil particles. Next, roots were depigmented with a 10% KOH solution. After incubation at 80°C for 30 min, the solution was poured off and replaced by a 1% HCl solution with a rinsing step in between. Roots were acidified for 20 min at room temperature before adding droplets of Parker Quink blue ink (2 droplets per 25 ml). This mixture was incubated at 80°C for 5 min and at room temperature for an additional 30 min. Finally, the staining solution was removed, roots were rinsed with water and an acidified glycerol solution (700 ml glycerol + 230 ml water + 70 ml 1% HCl) was added to extract the ink stain from the plant cells. Stained roots were observed microscopically with a Motic SMZ-168 stereo microscope (Motic, Causeway Bay, Hong Kong) and an Olympus BX41 light microscope (Olympus America, Inc., Center Valley, PA, USA) for presence of endophytic fungi.
8
Collagen Production Quantification in Renal Fibroblasts
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Collagen production was measured using a picrosirius red-based assay, as previously described [22 (link)]. Briefly, renal fibroblasts were treated with TGF-β1, AA2P, TGF-β1, and AA2P, or TGF-β1 and AA2P and 4μ8c. After 72 h, cells were fixed in methanol for 10 mins at -20 °C. Cells were subsequently washed and incubated with picrosirius red stain (0.1% Direct Red 80 in saturated picric acid; 1 h at room temperature). Cells were then washed 3× with 0.1% acetic acid and imaged using a BX41 Olympus light microscope. The dye was subsequently eluted from the cells with 0.1 N NaOH (10 min on a rocking platform) and moved to a 96-well plate. The absorbance of picrosirius red stain was measured using a Spectramax Plus Microplate Reader at 540 nm.
9
Chromogranin Immunohistochemistry in Epithelium
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Sections were examined under the BX41 Olympus light microscope. Images captured from four different fields containing squamous epithelium by the aid of the DP12 camera attached to the microscope and navigated by AnalySIS LS Starter software. Areas that were stained brown with chromogranin, were scored based on the intensity of cytoplasmic or nuclear staining and the percentage of the positive cytoplasmic area from the average of four microscopic fields at a fixed magnification of ×400. Nuclear staining was recorded by averaging the number of positive nuclei in four microscopic fields at a fixed magnification of ×400. Cytoplasmic and nucleus staining was graded based on the intensity (0, +1, +2, +3).
10
Histological Analysis of Liver Tissue
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Liver tissue samples were fixed in 4% paraformaldehyde, embedded in paraffin blocks and routinely processed. Sections cut at 4–6 µm were stained with hematoxylin/eosin and examined with an Olympus BX41 light microscope.
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