First true leaves of 14 DAG in vitro grown plants were cleared for 3 days in a solution prepared by mixing 120 g chloral hydrate, 7.5 ml glycerol and 150 ml of water, embedded in the same solution and imaged using the transmission light microscope Olympus Provis AX70. Trichome branch number was counted under transmission light microscope Olympus Provis AX70 with a 10 × objective, and branch length was measured after manual tracing on photos using the length measurement tool of ImageJ. All measurements were done in three biological replicates (at least 170 analyzed trichomes per genotype from one true leaf of at least 20 plants). Photos of cleared trichomes were captured using polarized light microscopy on Olympus AX70 Provis with a 10 × objective.
Provis ax70
Manufactured by Olympus
Sourced in Japan, France, Germany, United States, Poland
The Provis AX70 is an upright microscope designed for a variety of laboratory applications. It features a sturdy frame, coaxial coarse and fine focusing controls, and a trinocular observation tube. The Provis AX70 supports a range of objectives, from low to high magnification, to accommodate diverse sample types and observation needs.
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Lab products found in correlation
Dp manager imaging software, by Olympus (9 mentions)
Ikaros karyotyping software, by MetaSystems (6 mentions)
Dp30bw digital camera, by Olympus (5 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (4 mentions)
Vectashield, by Vector Laboratories (4 mentions)
Coolcube 1 b w digital camera, by MetaSystems (4 mentions)
Dp30w, by Olympus (3 mentions)
Microimage software, by Olympus (3 mentions)
Dp30bw, by Olympus (3 mentions)
Entellan, by Merck Group (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Dp30bw ccd camera, by Olympus (2 mentions)
Formalin, by Merck Group (2 mentions)
Application suite x 2, by Leica (2 mentions)
Alexa fluor 488 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions)
4 6 diamino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Biotinylated lectin antibody, by Vector Laboratories (1 mentions)
Infinity 2 3c, by Teledyne (1 mentions)
Image pro, by Media Cybernetics (1 mentions)
105 protocols using provis ax70
1
Trichome Branching Analysis in Vitro
2
Immunohistochemical Labeling of Calretinin and Tyrosine Hydroxylase in Teleosts
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-Polyclonal anti-calretinin (CalR) antibody 7696 (# 6B3 Swant, Bellinzona, Switzerland) at a 1∶10,000 concentration. This antibody has been widely used in the study of the neuroanatomy of teleosts, in adult animals as well as in embryos, larvae and juveniles [28] (link), [29] (link). Secondary antibody labeled with Cy3 (red) [30] (link).
-Anti-tyrosine hydroxylase (TH) antibody (Incstar, Stillwater, MN, USA) [31] (link), at a 1∶1,000 concentration. Secondary antibody labeled with Cy2 (green).
The sections were examined under a microscope (Olympus Provis AX70) coupled to a digital camera (XM10, Olympus). The images were coded green (Cy2) and red (Cy3), giving yellow co-localization in merged images. The images were adjusted for brightness, contrast and colors using Adobe Photoshop 7.0 (Adobe Systems) [27] .
-Anti-tyrosine hydroxylase (TH) antibody (Incstar, Stillwater, MN, USA) [31] (link), at a 1∶1,000 concentration. Secondary antibody labeled with Cy2 (green).
The sections were examined under a microscope (Olympus Provis AX70) coupled to a digital camera (XM10, Olympus). The images were coded green (Cy2) and red (Cy3), giving yellow co-localization in merged images. The images were adjusted for brightness, contrast and colors using Adobe Photoshop 7.0 (Adobe Systems) [27] .
3
Brain Section Immunofluorescence in APOE Mice
4
Microscopic Analysis of Plant Samples
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The samples fixed with FAA were subjected to the following treatments: (a) a part of the samples was embedded in paraffin [66 (link)]; (b) another part of the samples was embedded in LR-White® Hard Grade resin (London Resin Company, London, UK), according to Barreno et al. [30 ]; (c) finally, other samples were saved for observation using a low-temperature scanning microscope (LTSEM).
All the sections were observed with an OLYMPUS Provis AX 70 (Olympus, Hamburg, Germany) light microscope equipped with epifluorescence equipment. The observations with fluorescence and autofluorescence were performed with a UMWU fluorescence cube (excitation filter: 330–385 nm; barrier filter: 420 nm; dichroic mirror: 400 nm). All the images were photographed with a Lumenera Infinity 2-3C digital CCD colour camera (2080 × 1536 resolution, Lumenera, Ottawa, ON, Canada), captured with the Infinity Analyze® 7 v program. 7.1.0, and subsequently processed with the Photoshop CC® 2018 program, at the Jardí Botànic of the University of Valencia.
All the sections were observed with an OLYMPUS Provis AX 70 (Olympus, Hamburg, Germany) light microscope equipped with epifluorescence equipment. The observations with fluorescence and autofluorescence were performed with a UMWU fluorescence cube (excitation filter: 330–385 nm; barrier filter: 420 nm; dichroic mirror: 400 nm). All the images were photographed with a Lumenera Infinity 2-3C digital CCD colour camera (2080 × 1536 resolution, Lumenera, Ottawa, ON, Canada), captured with the Infinity Analyze® 7 v program. 7.1.0, and subsequently processed with the Photoshop CC® 2018 program, at the Jardí Botànic of the University of Valencia.
5
Hematoxylin-Eosin Staining Protocol
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Histological sections were obtained as mentioned above and stained with hematoxylin-eosin to observe possible morphological changes. Briefly, the technique involves immersing the sections in eosin for 1 minute, then washing with water every 30 minutes and further incubating for 1 minute in hematoxylin. Finally, the samples were dehydrated in ethanol of increasing concentration for 5 minutes each, ending with three tanks of xylene, for 3 minutes each. The slides were mounted in Entellan (Merck KGaA, Darmstadt, Germany) for analysis and storage. Images of hematoxylin-eosin staining were taken in a light microscope (Olympus Provis AX70) coupled to a digital camera (DP70, Olympus).
Finally, to adjust the brightness and contrast to those observed directly under the microscope, Adobe ® Photoshop CS2 ® version 9.0 (Adobe Systems) was used [27] .
Finally, to adjust the brightness and contrast to those observed directly under the microscope, Adobe ® Photoshop CS2 ® version 9.0 (Adobe Systems) was used [27] .
6
Lung Tissue Histomorphometric Analysis
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Conventional light microscopic examination of the lung tissue was performed using paraffin-embedded samples: 5 mm sections were stained with hematoxylin and eosin, and assessed in a blinded manner. The microscopical images were captured using an automatic microscope, Provis AX-70 with a camera (Olympus Optical, Tokyo, Japan). Morphometric analyses of the lung samples were performed with NIH 1200 image analysis software as previously described (12 (link)).
7
BV2 Cell Immunofluorescence Assay
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BV2 cells were seeded on Lab-Tek II chamber slides and then treated with linderone or LPS. Formalin was used for fixing cells. Next, the p65 antibody and Alexa Fluor 488 were treated. DAPI was used to visualize nucleifor. Finally, cells were observed and photographed using a fluorescence microscope (Provis AX70; Olympus Optical Co., Tokyo, Japan).
8
Measuring Intracellular Oxidative Stress
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HT22 cells were incubated with DCFH-DA (10 μM) for 20 min. After PBS washing, cells were observed and photographed under a fluorescence microscope (Provis AX70; Olympus Optical Co., Tokyo, Japan).
9
Immunofluorescence Imaging of BV2 Cells
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BV2 cells were cultured using Lab-Tek II chamber slides and then treated with S. horneri extract and its fractions (100 μg/mL) for 3 h before stimulation by LPS (1 μg/mL) for 1 h. Cells were fixed with formalin and then permeated with cold acetone. Next, cells were irradiated with p65 antibody and then incubated with fluorescein isothiocyanate (FITC)-labeled secondary antibody (Alexa Fluor 488, Invitrogen, Carlsbad, CA, USA). For nuclear staining, cells were treated with 1 μg/mL of 4′,6-diamidino-2-phenylindole (DAPI) for 30 min. Vectashield (Vector Laboratories, Burlingame, CA, USA) was treated after washing with PBS. Pictures of stained cells were confirmed using a Zeiss fluorescence microscope (Provis AX70; Olympus Optical Co., Tokyo, Japan) [30 (link)].
10
Immunofluorescence Analysis of NF-κB Activation
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BV2 and primary rat microglial cells were cultured on Lab-Tek II chamber slides and treated as described in the figure legends. The cells were treated with 80.0 μM steppogenin for 1 h, fixed in formalin, permeabilized with cold acetone, and then probed with a primary antibody against NF-κB and a fluorescein Isothiocyanate (FITC)-labeled secondary antibody (Alexa Fluor 488; Invitrogen, Carlsbad, CA, USA). To visualize the nuclei, the cells were treated with DAPI (1 μg/mL) for 30 min, washed with PBS for 5 min, and treated with 50 μL VectaShield (Vector Laboratories, Burlingame, CA, USA). The stained cells were visualized and photographed by using a Zeiss fluorescence microscope (Provis AX70; Olympus Optical Co., Tokyo, Japan) [38 (link)].
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