Maximum number of magnifications for a microscope

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Discussion Overview

The discussion centers around the maximum number of magnifications achieved by microscopes of various classes, with a focus on understanding the relevant properties of magnification and resolution in microscopy. Participants explore theoretical limits and practical applications in microscopy, including light and electron microscopy.

Discussion Character

  • Exploratory
  • Technical explanation
  • Conceptual clarification

Main Points Raised

  • One participant seeks concrete numbers regarding the maximum magnifications achieved by microscopes.
  • Another participant emphasizes that resolution, rather than magnification, is the critical property in microscopy, noting that visible light can resolve structures down to approximately 200 nm.
  • It is mentioned that electron microscopes can achieve resolutions down to the level of individual atoms, which suggests a different approach to magnification.
  • A participant explains the concept of diffraction-limited microscopy and introduces "superresolution microscopy" techniques that can surpass traditional limits, detailing two main categories: near-field probes and far-field imaging with localization techniques.
  • One participant reiterates the initial query about maximum magnifications and provides a rule of thumb for useful magnification in light microscopes and electron microscopes, linking it to numerical aperture and accelerating voltage, respectively.

Areas of Agreement / Disagreement

Participants express differing views on the relevance of magnification versus resolution in microscopy. While some focus on the maximum magnifications, others argue that resolution is the more significant factor, leading to an unresolved discussion on the topic.

Contextual Notes

Participants reference various microscopy techniques and their limitations, including the dependence on light wavelength and the complexities of near-field versus far-field imaging. There is an acknowledgment of the evolving nature of microscopy technology and its implications for magnification and resolution.

meteor
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Don't know where to post this, I hope is in the right place. What's the maximum number of magnifications that a microscope (of any class) has achieved to date? I want concrete ciphers.
 
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"Number" as in different magnification steps? You can magnify an image as much as you want, that is not the relevant quantity. The resolution is the interesting property, it is generally limited by the wavelength of the light or particle used to make the image: Visible light won't resolve structures smaller than ~200 nm, UV can resolve smaller structures but is likely to destroy chemical bonds. Electron microscopes can image individual atoms.
 
As @mfb mentioned, the important quantity in microscopy is resolution. Standard microscopy is diffraction-limited, meaning that the smallest resolved feature is on the order of the wavelength. In the case of visible light, this is about 200 nm. In the case of electrons, this can be angstroms.

However, the diffraction limit is a far-field effect (where the distance between the sample to image and the objective is large compared to the wavelength), and there are several techniques in existence nowadays for going beyond it ("superresolution microscopy"). These generally fall into 2 camps (below). The resolution for each is on the order of single nanometers when using visible light as the source.

The first is to use a near-field probe, where the distance between sample and objective is on the order of the wavelength or less. This is an active field of research and has only recently become feasible, mainly because Maxwell's equations in the near-field are much more complicated than in the far field (the desired information is contained in non-propagating evanescent surface waves), and their solution requires a fair degree of computational power.

The second technique is to use far field imaging along with some clever way of localizing light sources. The two most famous examples of this technique--awarded the Nobel Prize in Chemistry in 2014--are PALM (photoactivated localization microscopy) and STED (stimulated emission depletion). The techniques differ in their details, but both are based on the idea that a fluorescing molecule will display its light according to a predictable point spread function. If the point spread functions of two closely (sub-diffraction limit) spaced fluorophores can be distinguished, their most likely positions can be computed and an image can be built up. PALM does this by using different fluorophores of different colors, while STED does this by using partially overlapping lasers: one to excite a group of fluorophores and one to de-excite part of that group.
 
meteor said:
Don't know where to post this, I hope is in the right place. What's the maximum number of magnifications that a microscope (of any class) has achieved to date? I want concrete ciphers.

As a rule of thumb, the maximum 'useful magnification' in light microscopes under standard widefield conditions is approximately 1000 x (numerical aperture). For electron microscopes, the accelerating voltage is used instead of numerical aperture, and from what little I understand, the maximum useful magnification is about 1000 x (accelerating voltage)
 
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