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Microscopy

aysegulaysegul Posts: 33
in Sample Imaging
Hi everybody,
I wanted also to an important point about imaging and the need to find alternative imaging technique. Which is the best compatible microscope for Clarity? (Light sheet microscopy (Single plane illumination), multi-photon laser microscopy, confocal microscopy, 2-photon laser microscopy). Is it possible to image the sample without confocal microscopy ? Which microscopy can we use for displaying 3D?
Thanks in advance for your feedback and suggestions.
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Comments

  • jberryjberry Posts: 5
    Has anyone tried to use deconvolution microscopy?
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  • gjonesgjones Posts: 12
    Yes, we've deconvolved all our images.
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  • gjonesgjones Posts: 12
    @aysegul: that's a really broad question and therefore difficult to answer. If you just want a microscope for imaging Clarity samples then go for a basic confocal setup. Multiphoton has a range of uses and while the deeper light penetration advantage of multiphoton seems appealing, I would argue that Clarity does away with that necessity. Spend the money instead on nice objectives with a huge working distance such as the two Scale objectives (4 and 8mm working distances) that olympus have manufactured. We have a confocal microscope and a multiphoton and my preference is thus far the confocal setup as opposed to the multiphoton. Sorry about the brevity of this response, if you have any more specific questions then message me :)
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  • aysegulaysegul Posts: 33
    @gjones thank you for your enlightening explanation. Nobody explain this issue like that. If i have any more question i will consult you anymore :D
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  • aysegulaysegul Posts: 33
    @gjones i have a question again. Are these Scale objectives suitable for multiphoton microscope or confocal microscope ?
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  • jberryjberry Posts: 5
    Sorry, I meant taking image stacks with a widefield (ie non-confocal) epifluorescence scope and then deconvolving them. Has anyone tried this?
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  • gjonesgjones Posts: 12
    @aysegul: Yes, the Scale objectives can be used for multiphoton microscopy. They can also be used for confocal microscopy, depending on the type of confocal microscope. The objectives are quite long and therefore might have some trouble fitting between the stage and the part of the microscope where the objectives screw in. I would measure that distance on your confocal before you make a purchase. Remember, if the working distance is 4mm then you will be seeing your sample first from 4mm away, meaning the minimum distance you require is the length of the objective + ~4mm.
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  • MRGneuroMRGneuro Posts: 5
    @gjones was it these objective that you used; XLPN25XSVMP and XLSLPLN25XSVMP?
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