Fluorescent microscope as plate reader?

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Fluorescent microscope as plate reader?

Postby equibox » Feb 08 2013 4:45 pm

Hi all,

I have been wondering if it is possible to use a widefield fluorescence inverted microscope as a (96-well plate reader. I think with a low magnification lens and and an xy motorized stage it should be possible to read fluorescence intensity (or count cells) from the bottom of the plate. I googled on this topic but found nothing. Any suggestions are welcome.


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Re: Fluorescent microscope as plate reader?

Postby brianrasnow » Feb 27 2013 1:48 am

Lumascope (http://www.bulldog-bio.com/lumascope.html) might work for you as the simplest, smallest, fluorescent microscope plate reader, around $10k. I think the key challenge is where do you want to compromise in a plate reader, as everything has a price. Do you care to resolve cells across the entire well? Are you confident there is not significant non-specific fluorescence (that more detailed morphology might reveal). Will anyone review the morphology? Etaluma's stage isn't the fastest or most accurate to stitch multiple images together in each well, but it will take a sharp 1-3 megapixel picture in each well. Maybe you want to describe your application and images in more detail.
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Re: Fluorescent microscope as plate reader?

Postby rjm1975 » Aug 20 2014 3:39 am

I think it works in a pinch, as you say if you have an automated XY stage. With a really low magnification lens like a 1.25X you would be able to image the whole well. You would worry about whether you get even illumination over the whole well - a function of how well your light source is aligned. You will probably have less dynamic range and higher background with a CCD camera than you would with a microplate reader with PMT detector. It is probably also slower than a plate reader, especially if you want to do a multi-wavelength experiment. Maybe the reason you get no hits when you google it, is that most labs that are well equipped enough to have an automated XY stage, also have a plate reader?
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