All carotid arteries were collected from the origin of the common carotid artery to the bifurcation. The volumes of the neointima of different arteries were normalised by dividing by the longest carotid artery and multiplying by their lengths. Three-dimensional images of cleared carotid arteries were acquired using a light-sheet fluorescence microscope (Ultramicroscope II, LaVision Biotec, Bielefeld, Germany). We used a × 2.5 objective lens (Mv PLAPO 2VC, Olympus) or a × 4 objective lens (Mv PLAPO 2VC, Olympus) covered with a 6-mm working distance dipping cap. The laser source was a white light supercontinuum laser (SuperK EXTREME, NKT Photonics, Cologne, Germany). To observe cell distribution and artery morphology, the filters were set as 551/40 nm excitation and 567/50 nm emission for PKH26 and 640/30 nm excitation and 690/50 nm emission for Evans blue. The step size, scanning range, and exposure time were set to 5 μm, 1 mm, and 100 ms, respectively. 3D projections of the tagged image file format images of the artery were obtained using Imaris software (Bitplane, Oxford Instruments Company).
Mv plapo 2vc
Manufactured by Olympus
Sourced in Germany
The Mv PLAPO 2VC is a high-performance microscope objective lens designed for use in biological research and clinical applications. It features a Plan-Apochromat optical design with a numerical aperture of 1.4, ensuring excellent image quality and resolution. The lens is optimized for use with cover slips and provides a working distance of 0.13 mm.
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Lab products found in correlation
5 protocols using mv plapo 2vc
1
Volumetric Analysis of Carotid Arteries
2
Light-sheet Fluorescence Microscopy of Arteries
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Arteries were observed under the light-sheet fluorescence microscope (LaVision Biotec, Bielefeld, Germany). We used a 2.5x objective lens (Mv PLAPO 2VC, Olympus) or a 4x objective lens (Mv PLAPO 2VC, Olympus) covered with a 6mm working distance dipping cap. A supercontinuum white light laser (SuperK EXTREME 80 mHz VIS with wavelength from 400 to 2400nm, NKT Photonics, Cologne, Germany) was chosen as a laser source. To observe the cell distribution and artery morphology, the filters were set as 551/40nm excitation and 567/50nm emission for PKH26 and 640/30nm excitation and 690/50nm emission for Evans blue. The step size was set to 5µm. Carotid artery scanning range was set to 1mm. 3D projections of the tagged image file format (TIFF) images of the artery were obtained by using Imaris software (Bitplane, Oxford Instruments Company).
3
Light-Sheet Microscopy Imaging of Arteries
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Light-sheet microscopy (LSM) imaging was performed using a light-sheet microscope (Ultramicroscope II, LaVision Biotec, Bielefeld, Germany) equipped with a sCMOS camera (Andor Neo, Oxford Instruments Group), a 2× objective lens (MVPLAPO 2VC, Olympus) and a 6-mm working distance dipping cap. Cleared agar-artery samples were placed in a plastic holder and submerged in a reservoir filled with benzyl alcohol and benzyl benzoate (1:2). Arteries were imaged along two axes: the longitudinal (x-y) axis at 4× magnification (2× zoom), using a numerical aperture (NA) of 0.046 and a light sheet width of 1209; the transverse (x-z) axis at 10× magnification (5× zoom), using an NA of 0.046 and a light sheet width of 726. Two channels were imaged per sample: FITC-dextran stained channel imaged using 470/40 nm excitation and 525/50 nm emission filter; autofluorescence channel imaged using 710/75 nm excitation and 810/90 nm emission filter. The step size and exposure time were set to 5 μm and 100 ms, respectively. Images were acquired with the ImspectorPro software (LaVision BioTec, Bielefeld, Germany).
4
Colon Imaging with Cleared Tissue LSFM
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The cleared colons were scanned with a commercial LSFM (LaVision BioTec, Germany). We combined a magnification of with a objective lens (Mv PLAPO 2VC; Olympus) covered with a 6-mm working distance dipping cap. We used a supercontinuum white-light laser (SuperK EXTREME 80 MHz VIS with wavelength from 400 to 2400 nm; NKT Photonics, Cologne, Germany) as a laser source. The filters were set as excitation and emission for acriflavine and excitation and emission for Evans blue for the detection of cell morphology and vessels in the samples. The step size was set to and a total range of up to 2 mm for colon transversal scanning. The measurements were performed with 385-ms exposure times per slice, and a total imaging time of per colon sample. Imaris software (Bitplane, Oxford Instruments Company) was used to generate 3-D reconstructions of the tagged image file format images of the colons.
5
3D Imaging and Quantification of Atherosclerotic Plaque
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The samples were imaged by LSFM (Ultramicroscope II, LaVision Biotec, Bielefeld, Germany) with a 2.5 × objective lens (Mv PLAPO 2VC, Olympus). Excitation wavelengths included 488, 568, and 647 nm. The scanning z-step size was 5 μm. The Imaris software (Bitplane, Oxford Instruments Company) was utilized to perform sectioning and reconstruct three-dimensional (3D) projections of the arteries. To calculate the mean fluorescence value, the region of interest (ROI) of the plaque was first extracted by the threshold segmentation method. The sum of fluorescence intensity in the ROI of the plaque was then divided by the area of the plaque to determine the mean fluorescence value. The LSFM images were processed using the FIJI software. The ROIs of the plaque for both the PLXND1 and iNOS channels were determined to evaluate co-localization via Pearson's correlation coefficient (PCC) and Mander's overlap coefficient (MOC) of the plaque using the co-localization Finder plugin in FIJI.
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