When doing fluorescent microscopy, background is an issue. To reduce background,

Question

When doing fluorescent microscopy, background is an issue. To reduce background, a technique was

When doing fluorescent microscopy, background is an issue. To reduce background, a technique was devised to utilize the total internal reflection mechanism for the laser. It works by translating a laser be a m parallel to the principle axis so that it is totally internally reflected within the coverslip. A common setup includes a HeNe laser (633nm) with the objective, immersion oil and coverslip sharing an index of refraction of n=1.52. Water has an index of refraction n = 1.33. The objective has a focal length of f=0.30mm, the coverslip has a thickness of 170?m and the immersion oil's thickness is variable, but 100?m is common. Draw a ray diagram for this setup. Clearly label angles of interest Find d, the displacement from the principle axis to totally internally reflect the laser.

Explanation / Answer

b)d=r2-r1

1.52-1.33

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