For the visual observation of the distribution of live and dead bacteria in biofilm, LIVE/DEAD BacLight (Invitrogen Molecular Probes, Eugene, OR, USA) was used. SYTO9, which stains live bacteria with green fluorescence, and propidium iodide, which stains dead bacteria with red fluorescence, were mixed 1:1 and combined with PBS by adding 3 μL of the mixture per 1 mL of PBS to prepare a stain solution. The plates of the investigated materials were placed in the stain solution and reacted for 15 min. The stained biofilm or cells were observed under an epifluorescence microscope (Nikon, Eclipse 200).
Eclipse 200
Manufactured by Nikon
Sourced in Japan, United States, Argentina
The Eclipse 200 is a research-grade microscope designed for a variety of laboratory applications. It features high-quality optics and a stable, ergonomic design to support accurate and consistent observations.
Automatically generated - may contain errors
Lab products found in correlation
Rm2125, by Leica (3 mentions)
Dabco mounting medium, by Merck Group (2 mentions)
Live dead baclight, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Pegfp n1 tfeb, by Addgene (1 mentions)
Tissue tek, by Sakura Finetek (1 mentions)
Peroxidase conjugated avidin, by Agilent Technologies (1 mentions)
Biotinylated anti rabbit immunoglobulin, by Agilent Technologies (1 mentions)
Phalloidin tritc, by Merck Group (1 mentions)
Alexa fluor 555 goat anti rabbit igg, by Abcam (1 mentions)
Dm2500, by Leica (1 mentions)
Elements f3, by National Instruments (1 mentions)
Vectastain abc kit, by Vector Laboratories (1 mentions)
30 protocols using eclipse 200
1
Bacterial Biofilm Live/Dead Visualization
2
Overexpression of GFP-TFEB in AML12 Cells
3
Quantification of Lymphatic Vessel Density
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The quantification of LVD was performed as previously described by our group 9-11 (link, link, link). Briefly, stained TMA histologic sections were analyzed using standard light microscopy (Nikon, Eclipse 200). Under low magnification, the most vascularized intratumoral areas were identified. We counted the number of immunostained lymphatic vessels found in 10 “hot spot” areas at 400X magnification. The LVD for each case was expressed by the mean value (total number of vessels in 10 hot spot microscopic fields/10). The median of all the mean LVD values was the cutoff used to divide tumors into high or low LVD, as suggested by Hall et al. 12 (link).
4
Crayfish Intestine Histology Procedure
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The intestines of three crayfish in each treatment were used to histological observation. Midintestinal samples were quickly removed and fixed for 24 hr in 4% paraformaldehyde. After undergoing a dehydration process in ethanol with graded levels and hyalinization in xylol, the treated intestine was embedded in paraffin wax. The 5 μm tissue slices were stained with hematoxylin–eosin. The observation and photographs of paraffin sections were performed by a microscope (ECLIPSE 200, Nikon, Japan).
5
Histological Analysis of Ovarian Tissues
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The excised ovaries were fixed in paraformaldehyde (4%) for 24 h and then dehydrated in a gradient concentration of ethanol. The dehydrated ovarian tissues were then hyalinized in xylol and followed by embedding in paraffin. The embedded ovaries were sectioned with a rotary microtome at 5 μm thickness. Ovarian slices were stained with the hematoxylin and eosin (H&E). Ovarian stained sections were observed by an optical microscope (ECLIPSE 200, Nikon, Japan). Image-Pro Plus 6.0 software was used to take digital images, and oocyte (late-stage yolk synthesis stage oocyte and mature oocyte) number and diameter were statistically analyzed by ImageJ software.
6
Histological Evaluation of Tissue Architecture
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The full‐thickness sections were deparaffinized in xylene and re‐hydrated in descending ethanol series up to distilled water. The sections were stained with haematoxylin–eosin to evaluate the tissue architecture and cell infiltrate or with toluidine blue (TB) to evaluate the mucous secretion. For TB staining, the sections were soaked for 10 min in pre‐filtered 0.1% TB in 30% ethanol; washed in distilled water, dehydrated in ascending ethanol and mounted in synthetic resin. All the sections were stained in a single session to minimize artefactual staining differences. Digital images were acquired with a video camera‐equipped microscope (Eclipse 200; Nikon Instruments, Tokyo, Japan) with ×20 objective.
7
Histological Analysis of Shrimp Hepatopancreas
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Three shrimp were randomly selected from each replicate, and a total of 12 shrimp were selected from each group. The hepatopancreas tissue was immediately placed in centrifuge tubes containing 4% paraformaldehyde. After 24 hr of fixation, they were dehydrated, washed with chloroform, and embedded in solid wax blocks. The solid wax blocks were cut into 5 μm slices using a rotary microscope, stained with hematoxylin and eosin (H&E), observed, and photographed under an optical microscope (Eclipse 200, Nikon, Japan).
8
Immunostaining and Electron Microscopy of Liver
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Liver sections were fixed with 4% paraformaldehyde then incubated in 20% sucrose in PBS and embedded in optimal cutting temperature (OCT) solution at −20 °C. Liver cyrosections were immunostained with FoxO3a antibody followed by Dylight 549 conjugated secondary antibody and Hoechst 33342 staining. The images of sections were acquired under a Nikon Eclipse 200 fluorescence microscope with MetaMorph software. For electron microscopy (EM), liver sections were fixed with 2.5% glutaraldehyde in 0.1 mol/l sodium cacodylate buffer (pH 7.4), followed by 1% OsO4. After dehydration, thin sections were cut and stained with uranyl acetate and lead citrate. Digital images were obtained using a JEM 1016CX electron microscope.
9
Histological Analysis of Uteroplacental Units
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Uteroplacental units and maternal kidneys from rats at 6 days after injection were fixed for 48 h with formalin 10% in PBS 0.1 M (pH 7.4). Two or three uteroplacental units from every uterus (groups of 3–6 animals each) were randomly dissected, dehydrated, and embedded in paraffin. Sections of 5 μm were made by a microtome (Leica RM 2125, Wetzlar, Germany) and mounted on 2% silane coated slides. The sections were stained with hematoxylin and eosin (H&E) and observed by light microscopy (Nikon Eclipse 200, NY, USA).
10
Histological Examination of Intestinal Tissues
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One segment of ileum, cecum, and rectoanal junction (approximately 20 g each) from each animal was obtained at necropsy and immediately fixed in neutral buffered 10% formalin, dehydrated with alcohol, and embedded in paraffin. Sagittal cuts (5 µm) were made with a microtome (Leica RM 2125, Wetzlar, Germany), stained with hematoxylin and eosin (H&E), and mounted on 2% silane-coated slides. Tissue sections were observed by light microscopy (Nikon Eclipse 200, NY, USA).
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