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E200mv

Manufactured by Nikon
Sourced in Japan, China

The Nikon E200MV is a laboratory microscope designed for a wide range of applications. It features a binocular observation tube and a built-in LED illumination system. The E200MV offers a standard magnification range and is equipped with various objectives to accommodate different samples and research needs.

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13 protocols using e200mv

1

Protoplast Isolation from L-lysine-producing E. coli

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L-lysine-producing E. coli QDW, stored in glycerol, was streaked on an LB solid medium plate and cultured at 37 °C overnight in a constant temperature incubator. Single colonies were picked and transferred to an LB liquid medium in erlenmeyer flasks. The mixture was shaken at 37 °C for 24 h, and the bacterial concentration was recorded every 2 h. Finally, the growth curve of L-lysine-producing QDW was evaluated. In the middle and late period of the logarithmic growth phase of the bacteria, 10 mL of bacterial solution was taken and centrifuged at 2683.2 (× g) for 10 min at 4 °C. The bacteria were collected, and the SMM protoplast stable solution (pH = 6.5, 0.5 mol/L sucrose, 20 mmol/L anhydrous magnesium chloride, 0.02 mol/L maleic acid) was taken and centrifuged twice. The bacteria were suspended in 10 mL of SMM protoplast stable solution. The lysozyme solution was added and mixed well. The cells were incubated in a water bath at 37 °C, and the wall was broken by enzymatic hydrolysis. The separation process of protoplasts at 37 °C was observed under a microscope oil lens (E200MV, Nikon., Nanjing, China). Finally, the bacterial solution after enzymatic hydrolysis was centrifuged at 1509.3 (× g) for 15 min and washed once with SMM buffer; the protoplasts were then collected. The prepared protoplasts were stored in a refrigerator at 4 °C.
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2

Antifungal Efficacy of Streptomyces Extracts

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The inhibitory activities of Streptomyces extracts on spore germination of phytopathogenic fungi were observed using light microscopy (Nikon, E200MV, Japan). Each fungal strain was challenged in dose response to different extract concentrations (1/2 × EC50, 1 × EC50, or 2 × EC50). The mixture (0.1 mL) of extracts and fungal spore suspension (105 CFU/mL) at a ratio of 1:1 (v/v) was placed on a sterile glass slide and incubated in a moist chamber at 28°C for 20–24 h. For the control, we replaced extracts with sterile water. All experiments were performed in three replicates. We detected approximately 100 conidia in each field. The percentage of spore germination (PSG) was calculated as follows: PSG = (A – B)/A, where A and B represented the spore germination rate of control and treatment groups, respectively (Chen et al., 2018 (link)).
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3

Quantifying Airway and Alveolar Cell Positivity

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Conventional morphometric analysis was performed with the reticle of 100 points and 50 straight coupled to the microscope eyepiece (E200MV, Nikon Corporation, Tokyo, Japan) (Weibel, 2010 (link)). The total area of the reticulum was 104 μm2, which was fixed at the base of the epithelium and the positive cells were quantified. The positive cells at 1,000× magnification were counted in four airway fields, in at least three airways, and 10 random fields in the alveolar septa per animal. Based on the number of points that coincided with the positive cells within the lattice divided by the number of points coincident with the peribronchial area for the airways or alveolar walls (Leick-Maldonado et al., 2004 (link); Camargo et al., 2018 (link)).
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4

Quantifying Airway and Lung Tissue Structure

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The point-counting technique was employed using a reticle containing 50 lines and 100 points coupled to the eyepiece of a microscope (E200 MV, Nikon Corporation, Tokyo, Japan). The reticle had an area of 104 µm2. The airways were analyzed in four fields surrounding three airways per animal. In the lung parenchyma, ten random lung fields were analyzed. All analyses were performed at 1000× magnification. The number of positive cells was counted in the reticulum area as the number of reticulum dots in the tissue area. The number of positive cells per square area of tissue was obtained, with the results were expressed as positive cells/104 µm2 [42 (link)].
For picrosirius analysis (collagen fibers), images were captured under the Leica DM2500 microscope connected to a digital camera (Leica DFC420 Leica Microsystems, Wetzlar, Germany) using the Infinity Capture software (v 6.5.6, Lumenera Corporation, Ottawa, ON, Canada). The images were acquired using Image-Pro Plus 4.5 software (NIH, Bethesda, MD, USA), which allowed us to select from the spectrum to be developed. These shades represented the positive areas quantified in the previously determined area. We determined the volume fraction, expressed as a percentage of the total area of the frame [22 (link)].
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5

Plastic Decomposition in Yellow Mealworms

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The physical and chemical properties of yellow mealworms’ excrement after eating plastic alone and wheat bran control were tested by Phenom ProX scanning electron microscope (SEM, Thermo Fisher Scientific, Massachusetts, USA) and a specially designed and fully integrated apparatus, Energy Dispersive Spectrometer (EDS). Tests were carried out in the State Key Laboratory of Marine Resources Utilization in the South China Sea, Hainan University, China. Intestines stored in 10% buffered formalin were embedded in paraffin, cut at 5 μm, and stained with hematoxylin and eosin. These sections were then examined for histoarchitectural changes under an optical microscope (E200 MV, Nikon, Tokyo, Japan).
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6

Antifungal Activity of Extract against C. fragariae

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The stock solution (10 g l–1) was diluted into 5, 2.5, 1.25, 0.625, 0.313, 0.156, and 0.078 g l–1 using a double continuous dilution method (Wei et al., 2020 (link)). When 50 ml of the sterilized PDA liquid medium was precooled to 40–50°C, 1 ml of each extract diluent was added to generate different treatment concentrations (1.563, 3.125, 6.25, 12.5, 25, 50, 100, and 200 mg l–1, respectively). The inhibition ability of each extract concentration on the mycelial growth of C. fragariae was analyzed according to our previous method (Li X.J. et al., 2020 (link)). Antifungal activity was measured according to the growth diameter of C. fragariae at 28°C for 7 days. The morphological profile of pathogenic mycelia treated with 200 mg l–1 of extract was observed using an optical microscope (Nikon, E200MV, Japan). The half-maximal effective concentration (EC50) of the extract against C. fragariae was calculated in the light of a toxicity regression equation established by a least square method (Vanewijk and Hoekstra, 1993 (link)).
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7

Quantifying Alveolar Morphology via Mean Linear Intercept

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The mean linear intercept (Lm) is the most-used parameter to demonstrate the presence of alveolar enlargement [17 (link)]. The reticulum with 50 lines (100 points) was coupled under an optical microscope (E200 MV, Nikon Corporation, Tokyo, Japan), and the analysis was performed by counting the intersections between the reticulum lines and the lung parenchyma. A total of 20 distinct and random fields were analyzed in the lung parenchyma under 200× magnification on slides stained with hematoxylin and eosin. Lm was calculated using the equation: Lm = 2500 μm/number of times the line and the alveolar septum intersect [22 (link),43 (link),44 (link),45 ].
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8

Quantifying Alveolar Distension via Mean Linear Intercept

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Point counting was used to quantify the mean linear intercept (Lm) (Theodoro-Júnior et al., 2017 (link)). The Lm determines the index of the mean diameter of the distal air spaces, indicating the degree of alveolar distension (Margraf et al., 1991 (link); Hsia et al., 2010 (link)). The reticulum was attached to an optical microscope (E200Mv, Nikon Corporation, Tokyo, Japan), and the analysis was performed by counting the intersections between the reticulum lines and the lung parenchyma. Twenty distinct random fields of the lung parenchyma were counted at 200× magnification on slides stained with hematoxylin and eosin. Lm was calculated using the equation: Lm = 2,500 μm/number of times the line and the alveolar septum intersect.
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9

Alveolar Morphometry Analysis Protocol

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The Lm was measured using a microscope (E200MV, Nikon Corporation, Tokyo, Japan) with a 400× magnification and a reticulum with a known area (50 straight and 100 points) to calculate how many times the reticular lines intercepted the alveolar walls. The initial area of the alveolar walls was determined, excluding vessels and airways. For each animal, 20 lung parenchyma fields were randomly analyzed. Thus, the mean alveolar diameter was calculated according to the ratio of the area of the pulmonary parenchyma to the number of intersections between the lines and the alveolar walls. All Lm values were expressed in micrometers (24 (link)). We used this method of analysis to rule out any acute lung injury at the dose of LPS used in the present protocol.
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10

Foc TR4 Spore Germination Inhibition

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The spore germination of Foc TR4 was observed by a light microscope (Nikon, E200MV, Tokyo, Japan) after treatment with strain Y1-14 extracts. The spore suspension (1.0 × 106 CFU/mL) of Foc TR4 was fully mixed with the different concentrations of extracts (1 × EC50, 2 × EC50, 4 × EC50, and 8 × EC50). The mixture was dropped onto sterile glass slides and incubated at 28 °C for 12 h. The sterile water was used as a negative control. Spore germination was evaluated using the inhibition efficiency [29 (link)]. All experiments were performed in three biological replicates.
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