The skin samples remaining after the completion of the (FITC)-dextran transport studies were subject to fluorescence microscopy analysis. To perform the analysis the skin samples were prepared by cutting them in half along their diameter and embedding them in O.C.T. compound. The embedded samples were sectioned in 20 μm slices using a cryostat microtome (Bright Instruments, Huntingdon, UK). The skin sections were mounted, stained with DAPI and covered with glass cover slips. Fluorescence photomicrographs were obtained with a Zeiss Axioscope microscope equipped with a Nikon Digital Camera (DXM1200; Nikon, Kingston upon Thames, UK) at a magnification of 10 ×. Images were acquired using two fluorescence channels to allow the visualization of the cellular structures stained by DAPI (blue colour emission ~ 460 nm) and the FITC-dextran fluorescence signal (green colour emission ~ 520 nm). Tissue samples without FITC-dextran were also tested as controls. Images were processed using Image J Software (National Institutes of Health, Maryland, USA).
Axioscope microscope
Manufactured by Nikon
Sourced in United Kingdom
The Axioscope is a high-quality microscope designed for advanced laboratory applications. It features a stable and ergonomic design, delivering reliable performance and precise imaging capabilities. The Axioscope is equipped with a range of objective lenses, allowing for versatile magnification options to suit various research and analysis needs.
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Lab products found in correlation
4 protocols using axioscope microscope
1
Fluorescence Microscopy Analysis of FITC-Dextran Skin Penetration
2
Quantifying Prostate Cancer Nuclei Features
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The study material consisted of prostatectomy specimens from the files of Pathology Department. Immunohistochemistry for ERG was performed on tissue microarrays, as previously reported [3] . The cases were reevaluated and reclassified according to the current criteria [9, 10, 11] . From the obtained dataset four groups were established as a combination of the following features: lower-grade (Gleason pattern 3) or higher-grade (Gleason pattern 4) and ERG-or ERG+ (Fig. 1). The Table I shows the details of grading, however for making the analysis more evident, only Gleason pattern in the TMA core was used for analysis.
The images of hematoxylin-eosin stained tissue microarrays were taken on a Zeiss Axioscope microscope equipped with a 100× oil immersion lens using a Nikon D5100 digital camera. Pictures (Fig. 2) were transferred to a personal computer, converted from Nikon raw image format into TIF format and processed using color deconvolution algorithm. The resulting files were used for the segmentation of nuclei. The properly segmented nuclei were being selected by the operator until fifty nuclei were available for each case (Fig. 3). The images of the nuclei were then processed by a program which measured the geometric and textural features listed in Table II. Definitions of the form factors used:
The images of hematoxylin-eosin stained tissue microarrays were taken on a Zeiss Axioscope microscope equipped with a 100× oil immersion lens using a Nikon D5100 digital camera. Pictures (Fig. 2) were transferred to a personal computer, converted from Nikon raw image format into TIF format and processed using color deconvolution algorithm. The resulting files were used for the segmentation of nuclei. The properly segmented nuclei were being selected by the operator until fifty nuclei were available for each case (Fig. 3). The images of the nuclei were then processed by a program which measured the geometric and textural features listed in Table II. Definitions of the form factors used:
3
Microscopy Techniques for Imaging Biological Samples
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Light microscopy was performed with a Zeiss Axioscope microscope, equipped with a Nikon DXM1200F digital camera. Standard fluorescence microscopy was performed with an EVOS FL Auto Cell Imaging System (Life Technologies). Confocal laser scanning inverted microscopy was performed using a LEICA TCS SP8 system, equipped with an argon solid-state laser. The strains were imaged with an excitation 488 nm laser line and emission at 510–530 nm. For scanning electron microscopy (SEM), samples were fixed, for 4 h, with 5% (v/v) glutaraldehyde in 0.1 M phosphate buffer, pH = 7.2. The samples were then washed five times with the same buffer and dehydrated in a series of 25–100% ethanol washes. The fixed samples were dried for 1 h in a Critical Point Dryer (Quorum K850) and gold-palladium-coated in a Quorum SC7620 Spatter coater apparatus. The microphotographs were recorded using a scanning electron microscope JEOL model, IT-100 LV. The images were taken with an accelerating voltage of 20 kV, at high vacuum mode and secondary electron image.
4
Chiral Nematic CNC Suspension Characterization
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CNC suspensions at a range of concentrations were prepared by first concentrating the stock using a rotary evaporator (40 °C, pressure <40 mbar), then diluting to the desired final concentration using deionised water. Chiral nematic phase separation was observed in CNC suspensions at a range of concentrations. Glass capillaries were filled with the suspensions and sealed with UV epoxy (Norland Optical Adhesive 81), then left to equilibrate for at least 2 weeks. The anisotropic phase fraction was determined from the proportion of the suspension that appeared bright when viewed between crossed polarisers. The chiral nematic pitch of the anisotropic phase was measured as twice the periodicity of the fingerprint pattern observed in optical microscope images, which were collected using transmission illumination with crossed polarisers on a Zeiss Axioscope microscope equipped with a ×20 objective (Nikon T Plan SLWD, NA 0.3).
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