CytoViva-Admin

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  • #3339


    CytoViva-Admin

    Keymaster
    • ★★★★★

    Paula,

    The relative efficiencies of light scatter from different samples in fluid suspension using dark field illumination can be known by comparing the amplitude of the recorded spectrum at different wavelengths. To do this successfully the illumination cannot change between samples.

    If samples are on a solid background, efficiencies can be found by first illuminating a spectrally neutral target such as a spectralon ninety nine percent light reflectance standard. A dark field objective should be used so that only light scattered at an angle is recorded. The light guide will go to the back of the reflectance neck and connect with an adapter. The microscope is focused on the spectralon surface and spectrum is recorded. Then replace the spectralon with the surface containing the sample and scan again. Use the same exposure times for both recordings if possible, or adjust your measurement for the change in exposure time. This will show the sample’s light scatter efficiency versus wavelength.

    #3338


    CytoViva-Admin

    Keymaster
    • ★★★★★

    Paula,

    If you do not have the fluorescence module (DMF) to correlate nanoparticle spectral mapping to fluorescently labeled subcellular location, then you can use the Peak Location Classifier (PLC) tool in ENVI. This feature is under ‘CytoViva Analysis’ in the ENVI menu bar and will allow you to “map” pixels based on their peak wavelength. You will indicate your peak wavelength in nm and tolerance based on the fluorescent emission of your lysosome tracker. The noise floor may need to be raised if you notice that you are picking up noisy pixels.

    You will save this file which is a Classification file (similar to a SAM file) under Output Result to File. Hit the ‘double arrows’ (No Output Rule Image) and hit “OK”. Overlay this PLC Classification file the same way you would a SAM Classification. As long as the fluorescent tracker is detected with HSI, this is a quick way to locate your lysosomes. Then, you can build a spectral library of lysosomes based on the PLC results. Also create a nanoparticle spectral library and filter both against the negative control and against each other (careful with naming conventions here). Finally, you can map both libraries onto the datacube (see related thread on mapping 2+ spectral libraries).

    For related published papers, you can type “Cytoviva Lysotracker” or similar key words into Google Scholar.

    #3275


    CytoViva-Admin

    Keymaster
    • ★★★★★

    Hello, anchutka.
    On page 30 of the attached manual you will see instructions on Saving a Spectral Library and on page 37 you will see instructions on the steps for Spectral Angle Mapper.
    If this does not fully address your question please let us know.
    Thanks!

    #3251


    CytoViva-Admin

    Keymaster
    • ★★★★★

    Dr. Berthing,

    Thank you for the inquiry!
    Typically for the citation, researchers will state the location of our home office next to the initial reference of the equipment. For example:
    A CytoViva Enhanced Darkfield Hyperspectral Microscope system (Auburn, AL, USA) was used to…

    We can also help you describe the technology in your Methods section. Contact any of our technical team members and we can provide advice regarding what you should include based on your specific research purpose. If you do not have direct contact information for our technical team, please advise and we will have someone contact you directly.

    Alternatively, many researchers have chosen to include one of the CytoViva technical team members as a co-author of the paper. This is common when we have contributed heavily to the Methods and Discussion portions of the paper, as well as consulted in experimental design. We do not request this—it is entirely the researcher’s choice.

    Best regards,
    CytoViva

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