5. On a given microscope, the numerical apertures of the condenser and low power

Question

5. On a given microscope, the numerical apertures of the condenser and low power objective lenses are 1.25 and 0.25, respectively. You are supplied with a filter that selects a wavelength of 520nm.

a.) What is the limit of resolution on this microscope? Please be specific to how you came up with this answer, what formula did you use? b.) Will you be able to distinguish 2 points that are 300nm apart as being separate, or will they blur into one?

6. on the same microscope as in Question#5, the high dry objective lens has a numerical aperture of 0.85, what is the limit of resolution on this microscope, and will you be able to distinguish 2 points that are 300 nm apart as being separate, or will they blur into one? Explain answer.

Explanation / Answer

Answer

5a. Resolution R = 1.22 ?/ NA condenser + NA objective

R = 1.22 x 520/1.25 + 0.25

R = 634.4/1.5

R = 422.933 nm

5b. Since the resolution is 422.93 nm, the microscope can only distinguish objects greater than 422.93 nm apart and thus you will not be able to distinguish between the 2 points.

6. The given numerical aperture = 0.85

Resolution R = 1.22 x 520/(1.25 + 0.85)

R = 634.4/2.1

R = 302.1 nm

The microscope will not be able to distinguish the 2 points separated by 300 nm as it would become blurred.

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