Why is the Rayleigh Criterion not a definitive resolution limit for microscopes?

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The Rayleigh criterion provides a formula for microscope resolution based on the angular separation of two points and the aperture size, indicating that shorter wavelengths improve resolution. However, the criterion is not definitive, as it applies primarily to diffraction patterns rather than directly to object separation. The minimum angular separation for resolving two objects is equal to that for diffraction patterns, but this relationship is influenced by factors like signal-to-noise ratios and coherence of light. Additionally, the Rayleigh limit is specific to mutually incoherent points, which may not apply in all imaging scenarios. Ultimately, resolution limits are more nuanced than commonly perceived, with various criteria available for different contexts.
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From Rayleigh criterion, the resolution of microscope is given by

\sin\theta = 1.22\frac{\lambda}{D}

where D is the separation b/w two objects. Suppose D is constant, image and object distances are fixed, if we want to increase the resolution, we should increase the angle (theta) right? So we should use long wave-length light instead of short one? But in the text, they said short wavelength is preferred (in some case even use electron wave), why is that?
 
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Isn't D the aperture of the objective?
theta is the minimum separation between two points (on the object) - so you want to make theta as small as possible
 
mgb_phys said:
Isn't D the aperture of the objective?
theta is the minimum separation between two points (on the object) - so you want to make theta as small as possible

Oh! D is the separation between two points (for example, the distance between to pinholes) and I forget that theta estimates the size of zero-order disc of the diffraction pattern, so we should make it as small as possible. What was I thinking!? :(
 
KFC said:
Oh! D is the separation between two points (for example, the distance between to pinholes)
mgb_phys is correct. D is the apperture size. The formula that you present is for the diffraction limited resolution. θ is the minimum angular separation between two points that can be resolved.
 
turin said:
mgb_phys is correct. D is the apperture size. The formula that you present is for the diffraction limited resolution. θ is the minimum angular separation between two points that can be resolved.

Yes, D is the aperture size. Thanks for pointing that out. I still have a question. From Rayleigh's criterion, at small angle approxmiation, the limit of resolution gives

\theta = 1.22\frac{\lambda}{D}

where \theta gives the minimum angular separation of two patterns that can barely be resolved. That is, this criterion is applied on the diffraction pattern. But in the text, this also works on objects. Namely, the minimum angular separation of two objects that can barely be resolved is also equal to \theta = 1.22\frac{\lambda}{D}, why MINIMUM SEPARATION OF DIFFRACTION PATTERNS = MINIMUM SEPARATION OF OBJECTS?
 
Again, these criteria are based on heuristic arguments about how much signal-to-noise is required to differentiate an object from background. The Rayleigh criterion claims that neighboring airy discs can be separated if the central dip from the combined pattern is 20% below the maximum intensity. There are many resolution criteria- Sparrow's criterion is another. There are still more for pixelated detectors.

Even moreso, the Rayleigh limit is for two mutually incoherent points, something that may not be the case for a scene illuminated with partially coherent light.

The bottom line is that "maximum resolution" or "resolution limit" or all those other marketing buzzwords are not hard limits.
 
I do not have a good working knowledge of physics yet. I tried to piece this together but after researching this, I couldn’t figure out the correct laws of physics to combine to develop a formula to answer this question. Ex. 1 - A moving object impacts a static object at a constant velocity. Ex. 2 - A moving object impacts a static object at the same velocity but is accelerating at the moment of impact. Assuming the mass of the objects is the same and the velocity at the moment of impact...

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