On a particular microscope, the numerical apertures of the condenser and medium

Question

On a particular microscope, the numerical apertures of the condenser and medium power objective lenses are 1.25 and 0.25, respectively. You are supplied with a filter that selects a wavelength of 520nm. What is the limit of resolution on this microscope and will you be able to distinguish two points that are 330nm apart as being separate, or will they blur into one? Later on the same microscope, the high dry lens has a numerical aperture of 0.85. What is the limit of resolution and will you be able to distinguish two points that are 250nm apart as being separate, or will they blur into one?

Explanation / Answer

R=1.22/( NA of Cond. + NA of Obj.)...............Resoltion

R= 1.22*520 nm/(1.25+0.25)= 433.3nm

Since the microscope can only distingush between two points that are separeted by 433.3 nm and above. the microscope can't distinguish two points that are separated by a distance of 330 nm.

R= 1.22*520nm (1.25+0.85) = 302.1nm

So the microscope can't detect that the two objects are separated. Because the two objects that are separated by a distance fo 250nm looks blurred and appears as one object. Because the resolution power is greater than the separatin distance.

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