I understand specific techniques based on fluorescence and other methods can enhance the spatial resolution beyond the diffraction limit of lambda/2NA (xy-plane) in what we call as the super0resolution microscopy techniques. 1) But does a normal epi-fluorescence based visualization also award some sort of improvement to this? 2) Also let's say if I'm using a wavelength of 532nm and a 60x oil objective (NA=1.4) and I view 100nm sized gold nanoparticles, so my image would be spread out as per the PSF... but I'll still be able to view them right? So what will be the smallest size of particles that I can view with this system, 20 or 40 or 80nm?
Although you would not be able to resolve the object spatially to a spot smaller than the diffraction limit, you would be able to detect it if it is bright enough. The sensitivity depends on the detector, the signal-to-background ratio, how well the optical filters are matched to the fluorescence spectrum of the object, the quantum yield of its fluorescence, the number of fluorophores per particle, the rapidity with which the fluorescence becomes photobleached, and any quenching interactions of the fluorophores with their surroundings.
As Adam B Shapiro points out, being able to detect the FLUORESCENCE FROM a particle depends on the brightness of the fluoresence from the particle, not the size of the particle, and even single molecule detection has been explored (although the brightness may intrinsically depend on the particle size). Being able to detect should not be confused with being able to spatially resolve.
As Adam B Shapiro points out, being able to detect the FLUORESCENCE FROM a particle depends on the brightness of the fluoresence from the particle, not the size of the particle, and even single molecule detection has been explored (although the brightness may intrinsically depend on the particle size). Being able to detect should not be confused with being able to spatially resolve.
- Badges
- Science topic
- Similar topics
- Computer Science
- Images