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Powering Through It: Imaging Deep in Scattering Media

Powering Through It: Imaging Deep in Scattering Media

by Pina Colarusso and Craig Brideau | Jan 10, 2023 | Uncategorized

In the last post, we reviewed how scattering and absorption attenuate the intensity of the laser used to excite fluorescence and reduce the amount of emission signal that can make it to the detector(s). Tissue is very complex, and fluorescence from different dyes, as...
Friends and Foes: Scattering and Absorption

Friends and Foes: Scattering and Absorption

by Pina Colarusso and Craig Brideau | Nov 23, 2022 | Uncategorized

When carrying out fluorescence imaging in thicker samples, we often hear about how  “scattering” affects image quality by limiting the depth of imaging within the sample. Scattering occurs when light traveling through a sample encounters a change in refractive index, ...
Highlighting Color Correction

Highlighting Color Correction

by Pina Colarusso and Craig Brideau | Oct 20, 2022 | Uncategorized

Often we want to visualize multiple structures within the same preparation, and if the colors are not registered, this can lead to erroneous conclusions. For example, if you are counting nuclei, the color correction does not likely matter, but when examining whether a...
Making Sense of the Fine Details

Making Sense of the Fine Details

by Pina Colarusso and Craig Brideau | Sep 15, 2022 | Uncategorized

Over the next few posts, we will dig deeper into microscope objectives and explore how they can affect the quality of your imaging. Let’s start the series off with magnification and resolution, which can be a point of confusion when planning experiments.    Often,...
Photons to Pixels: Illuminating Bit Depth

Photons to Pixels: Illuminating Bit Depth

by Pina Colarusso and Craig Brideau | Aug 3, 2022 | Uncategorized

The meaning and relevance of bit depth is a common point of confusion for life scientists who use microscopes. Typically, when we mention bit depth in microscopy, we are referring to gradations in the intensity readings from a detector.  Let’s use a photomultiplier...
Two-Photon Microscopy: How to Minimize Optical Aberrations

Two-Photon Microscopy: How to Minimize Optical Aberrations

by Pina Colarusso and Craig Brideau | Jun 10, 2022 | Uncategorized

The main advantage of two-photon microscopy is the ability to image deeper into scattering samples, such as tissue and live specimens. One way that two-photon objectives assist deep imaging is by providing extended working distances compared to lenses for other...
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  • Powering Through It: Imaging Deep in Scattering Media
  • Friends and Foes: Scattering and Absorption
  • Highlighting Color Correction
  • Making Sense of the Fine Details
  • Photons to Pixels: Illuminating Bit Depth

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    Applications

    Applications

    Features

    Benefits

    Fixed cell imaging

    - Multi-channel acquisitionSPARQ can be used with any fluorophore

    Live cell imaging

    - Low laser intensity (comparable to standard widefield illumination)Reduces photobleaching and phototoxity, allowing for long imaging sessions
    - Fast 2D and 3D imaging With a frame rate of up to 10 FPS, catch your dynamic proccesses using SPARQ
    - Multi-dimensional acquisition: Time lapse, z-stacks, multi-channelsSet your usual multi-dimensional experiments and turn on SPARQ for optically sectioned images
    - Combinaison with transmitted light constrat methodsCombine your SPARQ images with any contrast modes, without even changing objectives

    Tissue slice imaging



    - High penetration depthSince the speckles remain sharp in depth, get the same optical sectioning capabilities as you go deeper
    - Modification of optical thickness Adjust SPARQ's thickness factor parameter to change the axial resolution without affecting the lateral resolution
    - 3D imaging and reconstructionCreate high quality 3D images by rejecting out-of-focus fluorescence

    Whole mount and cleared tissue imaging

    - Large area stitchingKeep the same imaging speed regardless of the field of view. SPARQ works with any camera
    - Use different objective magnifications Simply change objectives without the need to re-calibrate or to modify the SPARQ parameter
    - Works with any kind of objectivesUse high N.A, LWD or phase contrast objectives. It will not affect the SPARQ images
    - Keep high axial and lateral resolutionImprove the optical sectioning without sacrificing the lateral resolution as with standard confocal microscopy

    In vivo imaging (on 2P)

    - Can be used in combinaison with photo-stimulationUse the same visible lasers to photo-stimulate and do SPARQ imaging (different light paths)
    - Offers optical sectioning with visible lasers on a 2P optimized systemIncrease the flexibilty of your 2P system by adding SPARQ and get visible light confocality
    - Can be used in vivo for positioning and other fluorescence usesA one push button allows you to easily switch between SPARQ and other imaging modes
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