A student has set up an experiment using a confocal light microscope where they

Question

A student has set up an experiment using a confocal light microscope where they would like to count the number of individual proteins in some small area on substrate; each protein of interest is bound to a fluorescent tag and fluoresces at 550 nm, and is on the order of 125 nm in size. He asks your advice on what to expect. A) This should not be a problem so long as he limits the amount of light entering the detector. B) The results will be null, because 550 nm wavelength is not visible to the human eye. C) This experiment should work so long as he chooses a longer wavelength tag. D) The result will be misleading they will not be able to resolve individual proteins in the visible spectrum. E) It is not clear what the results might be; more information would be needed.

Explanation / Answer

In confocal light microscopy, resolution of the image increases with shorter wavelength.

So, 550 nm light can not produce resolvable image of proteins of size 125 nm.

Hence, Answer is (D) the results will be misleading, they will not be able to resolve individual proteins in the visible spectrum. ANS

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