How do optical aberrations affect microscopy axial resolution?

In summary, the conversation discusses the attempts to improve the axial resolution in microscopy, which is typically three times lower than the lateral resolution due to the shape of the lens' focal spot. This has been addressed through the use of additional lenses and techniques like 4Pi Fluorescence microscopy. However, there are still limitations due to factors such as spherical aberration. The conversation also touches on the difference between axial and lateral resolution and the use of confocal microscopy to improve resolution.
  • #1
kattie
2
0
It is actually not a homework, but rather a paper I need to present, talking about the advance attempts to raise microscopy axial resolution.
It is said the axial resolution is usually 3 times lower than lateral. The explanation is that the lens from 1 side of focus plane can only produce a segment of spherical wavefront, and thus, making the focal spot longer than wide.
(If there were a whole spherical wavefront, the focal spot would be spherical too.)
People also have tried to get close to this spherical wavefront by introducing another lens at the opposite side of the focus plan, thus reduce the length of the focal spot. (4Pi Fluorescence microscopy - invented by http://www.mpibpc.gwdg.de/groups/hell/4Pi.htm)

However, I don´t get the idea here. Why could a segment of spherical wavefront be responsible for the longer-than-wide focal spot?

By the way, my background is not in Physics so please make it as simple as possible. And sorry if there is a similar question already (I can´t find the search tab to look for it)

Thanks a lot.
 
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  • #2
I don't really know how you get a solid angle of 4π, but as far as your question goes, lenses are not perfect, and this is just something that happens.

So there are a lot of things that contribute to a non-perfect lens, such as spherical aberration (what you are talking about). Most of the effects, comas, airy effects, etc. can be reduced by using aspheric lenses and making everything as paraxial as possible, but at a certain point there is not much else you can do.

Phase contrast imaging is pretty good, and the resolution can get pretty close to the wavelength limit. For imaging at sub-wavelength limits, I have no idea how it works. What is your background in?
 
  • #3
I major in Biology, and that´s why we have to study about microscopy.
So now I should learn about "spherical abberation"? :D
 
  • #4
The axial and lateral resolution are different properties.

Focusing brings a bunch of rays together into an approximate spot - called the 'circle of least confusion' in optics. Objects slightly in front of and slightly behind the best focus are also almost in focus. This is the depth of field in photography - otherwise when you took a photograph only an infinitely thin layer would be in focus.
You try and design microscope lenses to have as small a depth of field as possible (a side effect is that this is also the design that collects the most light) but because the sample is slightly transparent an area infront of and behind will also be in focus.
The way to reduce this is confocal microscopy where you also use a lens to focus the illumination in a thin in-focus layer.

Lateral resolution is where two points on the image are just far enough apart not to be a single blur. Again this is due to optical aberations not bringing the light from slightly different directions into the same place = the 'circle of confusion' not being a perfect point.
This is reduced by reducing sphericla abberation and other geometric optical effects.
 

1. What is the focusing process in optics?

The focusing process in optics refers to the manipulation of light waves to converge or diverge at a specific point, creating a sharp image. This is achieved through the use of lenses or mirrors that bend or reflect light rays in a controlled manner.

2. How does the focusing process work?

The focusing process works by using the principle of refraction or reflection. When light passes through a lens, it is bent in a specific way, depending on the shape and curvature of the lens. This bending of light helps to bring all the rays to a single point, creating a focused image. Alternatively, a curved mirror can also reflect light rays to a focal point, achieving the same result.

3. What are the types of lenses used in the focusing process?

The two main types of lenses used in the focusing process are convex and concave lenses. Convex lenses are thicker in the middle and thinner at the edges, and they converge light rays to a focal point. Concave lenses, on the other hand, are thinner in the middle and thicker at the edges, and they diverge light rays, creating a virtual image.

4. What factors affect the focusing process?

The focusing process can be affected by several factors, including the curvature and shape of the lens or mirror, the distance between the lens and the object, and the wavelength of light. Additionally, the refractive index of the material the lens is made of and any imperfections in its surface can also impact the focusing process.

5. How is the focusing process used in everyday life?

The focusing process is used in many everyday devices, such as cameras, telescopes, and microscopes, to create sharp images. It is also used in corrective lenses for glasses and contact lenses to help people with vision impairments see clearly. In addition, the principles of focusing are essential in various medical procedures, such as laser eye surgery.

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