Lungs were dissected, inflated with 1–1.5mL of 2% low-melting agarose (Seaplaque, 50100) dissolved in PBS. Inflated lungs were placed on ice until agarose solidified. Whole lobes were stained with LEL Fluorescein (Vector Laboratories, FL-1171, 1:500) to visualize alveolar cups. Following media acclimation, whole lobes were imaged using an Olympus FVMPE-RS multiphoton laser scanning microscope with a 25x NA 1.05 water-immersion objective. The system has a Spectra Physics Insight X3, laser tunable from 680nm-1300nm and a fixed 1045nm laser. 880 nm excitation was used for LEL fluorescing imaging and 1045 nm laser was used to acquire tdTomato signal with a 2 μm z step-size. Images were acquired every 10 minutes for 3 hours.
Fvmpe rs multiphoton laser scanning microscope
Manufactured by Olympus
Sourced in Japan
The FVMPE-RS multiphoton laser scanning microscope is a high-performance imaging system designed for advanced biological research. It utilizes multiphoton excitation technology to enable deep tissue imaging with minimal photodamage. The core function of the FVMPE-RS is to provide researchers with a versatile and powerful tool for visualizing and analyzing complex biological samples.
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Lab products found in correlation
3 protocols using fvmpe rs multiphoton laser scanning microscope
1
Alveolar Structure Visualization via Multiphoton Microscopy
2
Multiphoton Imaging of Collagen Constructs
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Imaging was performed on whole-mount constructs in Opti-MEM medium without phenol red using the Olympus FVMPE-RS multiphoton laser scanning microscope with a water immersive objective at ×25. Acellular gels were imaged in parallel and used as background controls. Multiphoton excitation was performed with Dual Line Insight X3 laser at an excitation of 880 nm. Second-harmonic generation (SHG) and ScxGFP were recorded by GaAsP PMT detectors (4 G and 3 G, respectively). All parameters (ie, laser intensity, gain, high voltage, and offset) were selected to minimize background noise without oversaturation and were held constant for all constructs. Images were converted using the Bio-Formats plugin and batch-processed in ImageJ. Mean pixel intensity were analyzed to quantify SHG signal and collagen density.
3
3D Imaging of Bone Defects
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3D fluorescent images were acquired using a Nikon A1R confocal laser scanning microscope with a 20× objective lens. The z stacks of 80 μm in height and an x-y detect area at a size of 1,024 × 1,024 pixels, with a resolution of 0.624 μm, were taken for each slide. The 1-mm defect was imaged by tiling three z stacks, spanning 1,500 mm along the long axis of the tibia.
The second harmonic generation of collagen fibers was acquired on an Olympus FVMPE-RS Multiphoton laser scanning microscope (Japan). Images were excited with an 860-nm laser, and emissions were detected using 420–465 filters. The z stack of 40 μm height and an x-y detect area, at a size of 1,024 × 1,024 pixels, with a resolution of (0.623) μm, were taken for each sample.
The second harmonic generation of collagen fibers was acquired on an Olympus FVMPE-RS Multiphoton laser scanning microscope (Japan). Images were excited with an 860-nm laser, and emissions were detected using 420–465 filters. The z stack of 40 μm height and an x-y detect area, at a size of 1,024 × 1,024 pixels, with a resolution of (0.623) μm, were taken for each sample.
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