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Ti sapphire chameleon ultra laser

Manufactured by Coherent Inc
Sourced in United States

The Ti:Sapphire Chameleon Ultra laser is a tunable solid-state laser that operates in the near-infrared region of the electromagnetic spectrum. It utilizes a titanium-doped sapphire crystal as the active medium, which allows for a broad tuning range and the generation of ultrashort pulses.

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4 protocols using ti sapphire chameleon ultra laser

1

Collagen Fiber Orientation in Tendon

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Tendon sections (N=3) were imaged using a Zeiss 710 NLO inverted
confocal microscope (Carl Zeiss Microscopy) with a mode-locked Ti:Sapphire
Chameleon Ultra laser (Coherent Inc.) in combination with non-descanned
detection (NDD) to visualize the collagen fibers and quantify their orientation
and distribution as previously described31 (link). The laser was set to 800 nm and emission was filtered
from 380 - 430 nm. Second harmonic generation (SHG) images were collected using
a Plan-Apochromat 20x objective and Zeiss ZEN software. The fiber direction was
estimated using OrientationJ distribution, an ImageJ plug-in (NIH) developed for
directional analysis. A distribution of local angles was generated for each
optical slice, where 0° aligned to the horizontal axis (length-wise along
the tendon) and ± 90° to the vertical axis. The mean and standard
deviation of collagen fiber orientation was calculated from the distribution of
each image. For analysis, eight sections of supraspinatus tendon (each section
was imaged at both proximal and distal ends) were used.
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2

Visualizing Collagen Fiber Architecture in Heart Valve Leaflets

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Representative leaflet specimens were also subjected to second harmonic generation (SHG) imaging to visualize local collagen fiber architecture in the belly region of the leaflet, as for biaxial tensile testing. Leaflets were imaged on a Zeiss 710 NLO inverted confocal microscope (Carl Zeiss Microscopy, LLC, Thornwood, NY, USA) equipped with a mode-locked Ti:Sapphire Chameleon Ultra laser (Coherent Inc., Santa Clara, CA) in combination with non-descanned detection (NDD). The laser excitation was set to 800 nm and backward SHG signal was filtered from 380–430 nm. Samples were kept hydrated with saline solution during imaging to prevent drying artifacts and covered with #1.5 coverslips. SHG was collected from the atrial side of the leaflets using a Plan-Apochromat 63x oil immersion objective for correlation of local collagen fiber architecture to the local biaxially-derived mechanical properties. Image resolution was set to 0.44 × 0.44 μm2 per pixel at 8-bit pixel depth.
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3

Quantifying Collagen Fiber Structure

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To quantify the collagen fiber structure, tested biaxial samples from Caballero et al. (2017) [5 (link)] were imaged in the central region (delimited by the graphite markers) using second harmonic generation (SHG) imaging. Tissues were imaged on a Zeiss 710 NLO inverted confocal microscope (Carl Zeiss Microscopy, LLC, Thornwood, NY, USA) equipped with a mode-locked Ti:Sapphire Chameleon Ultra laser (Coherent Inc., Santa Clara, CA) in combination with non-descanned detection (NDD). The laser was set to 800 nm and emission was filtered from 380–430 nm [23 (link)]. Samples were kept hydrated with saline solution during imaging to prevent drying artifacts and covered with #1.5 coverslips. SHG was collected from the smooth side of the tissue using a Plan-Apochromat 40x oil immersion objective. Zeiss ZEN software was used to visualize and export image stacks for analysis. Image resolution was set to 0.35 × 0.35 μm2 per pixel at 12-bit pixel depth.
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4

Multiscale Characterization of Ligament Microstructure

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Upon completion of biaxial mechanical testing, the tissue samples were imaged using the SHG technique at the unloaded state. We utilized a Zeiss 710 NLO inverted confocal microscope (Carl Zeiss Microscopy, LLC, Thornwood, NY, USA), equipped with a mode-locked Ti:Sapphire Chameleon Ultra laser (Coherent Inc., Santa Clara, CA), a non-descanned detector (NDD), and a Plan-Apochromat 40× oil immersion objective. The laser was set to 800 nm and emission was filtered from 380–430 nm. Samples were kept hydrated with saline solution during imaging to prevent drying artifacts and covered with #1.5 coverslips. Samples were imaged inside the area delimited by the graphite markers, and 2D image slices were collected in the thickness direction from the smooth side of each sample. A 2D slice has 512×512 pixels to 1024×1024 pixels, and for each sample the number of slices was varied to cover the thickness. In total, we obtained 3D SHG images (size from 512×512×N to 1024×1024×N) of 48 tissue samples from different animal subjects, and the corresponding mechanical testing data. Representative SHG images of a GLBP sample are shown in Figure 2, with a total of 18 slices (N=18) through the thickness. It can be seen that geometry variation (e.g. fiber waviness) in each imaging plane is much larger than that in the thickness direction as shown in Figure 2.
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