FAQ's about microscopy


This is part of the Huygens Faq.

Full list


  • What's a bandlimited system? Answer.
  • How far above and below the in focus information should I sample? Answer.
  • What are the pros and cons of piezo-electric vs step motor focusing control? Answer.
  • Do I need to use all three channels when observing through a broad bandpass barrier filter? Answer.
  • What do you mean by the mounting medium? Answer.
  • Which pinhole size to use in case of mechanical hysteresis? Answer.
  • May I clip bright pixels which dominate the scene? Answer.
  • Nyquist rate - What's arcsin/asin? Answer.
  • How to compute the back projected pinhole radius? Answer.
  • Backprojected pinhole radius for Biorad confocals? Answer.
  • What does the message: 'total reflection at glass/medium interface (...)' mean? Answer.
  • I have heard that imaging diatoms is a difficult task, why? Answer.
  • Should PSFs be produced for different wavelength when working in the visible light range? Answer.
  • What emission wavelength to use to represent the detected spectrum? Answer.
  • What to use as excitation wavelength: laser line or fluorescence spectrum peak Answer.
  • What is the refractive index of the embedding medium Vectashield? Answer.
  • What is the refractive index of the medium? Answer.
  • Photon counting - why do people sometimes question its practicality? Answer.
  • A raw 1 micron bead looks very elongated along the optical axis. Why? Answer.
  • Which wavelength do I have to set for multi-photon excitation? Answer.
  • Which pinhole size to use in the Leica SP2 confocal microscope? Answer.
  • How to estimate the back projected pinhole value from the Airy Disk size? Answer.
  • When is the RI / NA combination correct? Answer.
  • How to calculate the X and Y sample sizes using a CCD camera. Answer.
  • Do you know of any suitable solid mountants with suitable refractive indices? Answer.
  • When deconvolving 2-photon data what microscope type should we choose? Answer
  • The PSF looks "banana-shaped" in the x-z or y-z planes. What could be the reason? Answer.
  • How do you measure the Numerical Aperture (NA) from the experimental PSF ? Answer.
  • How to immobilize a sample with agarose or gelatin Answer.
  • What are the characteristics of the Confocal, Widefield, Multiphoton (and other) modules? Answer.