Why Fluorescence is detected @right angle to the excitation?

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Discussion Overview

The discussion centers on the optimal angle for placing a detector relative to a light source when observing emission radiation from fluorescent materials. It explores the common practice of using a right angle for detection and the implications of alternative angles, particularly in the context of minimizing interference from reflected or transmitted light.

Discussion Character

  • Exploratory
  • Technical explanation
  • Debate/contested

Main Points Raised

  • One participant inquires about the rationale behind using a right angle for detector placement, referencing that it minimizes the risk of transmitted or reflected light reaching the detector.
  • Another participant notes that various materials reflect light at shallow angles, which could affect fluorescence measurements, although they do not provide specific details.
  • A participant explains that in right angled illumination, a half silvered mirror at 45° can combine the incident and emitted beams, but this setup results in a loss of intensity due to the nature of the optical paths involved.
  • The same participant suggests that right angled illumination simplifies engineering and avoids complications associated with Brewster's angle.
  • One participant mentions that wavelength filters can effectively block light from the initial source, indicating that this may mitigate some concerns about reflected light interference.
  • Another participant clarifies that the brightness of the full moon, while related to surface illumination, is not directly relevant to the fluorescence discussion, emphasizing that there are additional factors influencing brightness in different lunar phases.

Areas of Agreement / Disagreement

Participants express differing views on the implications of detector placement angles and the relevance of certain optical phenomena, indicating that multiple competing perspectives remain without a clear consensus.

Contextual Notes

Some assumptions about the behavior of light at different angles and the effectiveness of various optical components are not fully explored, leaving room for further investigation into the specifics of fluorescence detection.

Shi
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Hi everybody,
I would like to know what would be the best angle (or best geometry) to put the detector relative to the light source in order to observe the emission radiation from the fluorescent materials?
As I know the most common geometry used for fluorescence is right angle observation. Detector is placed at 90 deg to incident light to minimize the risk of transmitted or reflected incident light reaching the detector (from wiki). Why?!
If we place detector at 45 deg or at smaller angle to the source, what would be happened?
 
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Various materials tend to reflect under shallow angles, or directly back to the source. I don't know details about it in terms of fluorescence measurements, but that effect is well-known in astronomy.
 
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Shi said:
Detector is placed at 90 deg to incident light to minimize the risk of transmitted or reflected incident light reaching the detector (from wiki). Why?!
Near the sample, the incident beam and the emitted beam pass along the same optical path but in different directions. The source and the detector cannot be in the same place so the two beams must be combined or separated at some point.

For the right angled illumination case, a half silvered mirror mounted at 45° would combine the beams. But the half silvered mirror would reduce the detected beam to 25%, since half the illumination is lost on the first pass, before half the emission is lost on the second pass. Since the two beams have different wavelengths, a wavelength sensitive dichroic mirror is used to effectively brighten the response for the same illumination source.

I believe that right angled illumination with a 45° mirror is used because it leads to simple compact engineering and avoids the Brewster angle. https://en.wikipedia.org/wiki/Brewster's_angle

mfb said:
Various materials tend to reflect under shallow angles, or directly back to the source. I don't know details about it in terms of fluorescence measurements, but that effect is well-known in astronomy.
The astronomical phase curve is best explained by the relative brightness of our moon in different phases. A full moon has more surface illuminated by the Sun that is also visible from Earth. The new moon returns little light because we cannot see much of the illuminated surface. The shape of the curve is determined to some extent by surface texture. That cannot be related to the florescence microscope situation.
 
Baluncore said:
A full moon has more surface illuminated by the Sun that is also visible from Earth.
That is not the only effect, and it is not the effect I was discussing. The full moon is much brighter than you would expect based on illuminated area. The linked article explains those brightness excesses and opposition surges.
But I see that wavelength filters work fine to stop light from the initial light source, so this is not an issue here.
 

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