For high contrast images of fluorescently stained tissue, we used inverted Olympus IX83 microscope with a total internal reflection fluorescence objective (UAPON 100X/1.49NA, Olympus), sCMOS camera (ORCA_Flash4.0 V3, Hamamatsu), and 488nm excitation laser (Genesis MX488, Coherent). In order to ensure E-cadherin distribution was being quantified at the same region of the epithelium, tissue was evaluated initially under 10x magnification to identify mid-epithelial areas of flat tissue located close to the coverslip. We then chose sharply in focus parts of tissue areas and measured the full width at half maximum (FWHM) of E-cadherin clusters and the size of the non-fluorescent, dark core within an E-cadherin cluster.
Uapon 100x 1.49na
Manufactured by Olympus
Sourced in Austria
The UAPON 100X/1.49NA is an oil immersion objective lens for use in advanced microscopy applications. It features a numerical aperture of 1.49 and a magnification of 100X, providing high-resolution imaging capabilities. The lens is designed for use with oil immersion techniques to optimize optical performance.
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3 protocols using uapon 100x 1.49na
1
High-Contrast Fluorescence Imaging of Tissue
2
Dual-color Imaging of Platelet Ultrastructure
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Images were acquired using a modified Olympus IX81 inverted epifluorescence microscope with an oil-immersion objective (UApo N 100x/1.49 NA, Olympus, Vienna, Austria). The sample was positioned with nanometer precision on a XYZ piezo stage (P-733.3DD, Physical Instruments) on top of a mechanical stage with a range of 1 × 1 cm adjusted by precision screws (TAO, JPK Instruments, Berlin, Germany). A tube-lens with an additional magnification of 1.6 was used to achieve a final imaging magnification of 160 (corresponding to a pixel size of 100 nm). Platelets were illuminated with a 642 nm laser light from a diode laser (Omicron-laserage Laserprodukte GmbH, Phoxx 642, Rodgau-Dudenhofen, Germany), a 488 nm laser light from a solid-state laser (diode-pumped, Toptica Photonics, Graefelfing, Germany), and a 405 nm laser light from a diode laser (Insaneware, Gladbeck, Germany). The signal was detected using an Andor iXonEM+ 897 (back-illuminated) EMCCD camera (16 μm pixel size). The following filter sets were used: dichroic filter (ZT405/488/561/640rpc, Chroma, Olching, Germany), emission filter (446/523/600/677 nm BrightLine quad-band band-pass filter, Semrock, Rochester, NY, USA), and an additional emission filter (HQ 700/75 M, NC209774, Chroma Technology GmbH, Olching, Germany).
3
High-Contrast Fluorescence Imaging of Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For high contrast images of fluorescently stained tissue, we used inverted Olympus IX83 microscope with a total internal reflection fluorescence objective (UAPON 100X/1.49NA, Olympus), sCMOS camera (ORCA_Flash4.0 V3, Hamamatsu), and 488nm excitation laser (Genesis MX488, Coherent). In order to ensure E-cadherin distribution was being quantified at the same region of the epithelium, tissue was evaluated initially under 10x magnification to identify mid-epithelial areas of flat tissue located close to the coverslip. We then chose sharply in focus parts of tissue areas and measured the full width at half maximum (FWHM) of E-cadherin clusters and the size of the non-fluorescent, dark core within an E-cadherin cluster.
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