Structured Illumination Microscopy

SIM cut-off frequency, lateral resolution and raw image count for linear and saturated structured illumination microscopy.

Model: Textbook approximation

Results

Widefield cut-off 2NA/λem5.38 cycles/µm
Widefield resolution λem/(2NA)185.7 nm
Pattern period 1/k₀174.3 nm
SIM cut-off 2NA/λem + m·k₀11.12 cycles/µm
SIM resolution 1/k_c89.9 nm
Improvement over widefield2.07×
Raw images (3 angles × 3 phases)9

The pattern frequency lies beyond the detection cut-off: the stripes are not visible in the raw images, so the pattern phase and frequency are estimated from the overlap of the shifted passbands.

k₀ = p · 2NA/λex: two beams at the pupil edge give the finest pattern (p = 1). Linear SIM: k_c = 2NA/λem + k₀ (Gustafsson 2000).

Saturated SIM adds harmonics m·k₀ (Gustafsson 2005). Separating orders −m … m needs 2m + 1 phases per angle.

The resolution 1/k_c is the Abbe-type period, like λ/(2NA) for widefield. Reconstruction (Wiener filtering) and noise set how much of the support is usable.

Frequency support along one pattern direction

Each copy is drawn at unit height; in the raw data the first orders carry half the modulation depth, and harmonics less.

Model and references

Textbook approximation. A standard approximation from the literature (for example paraxial, plane-wave or Gaussian-beam), valid within the range the page states. About the tiers

Frequency-support model: the cut-off is the detection cut-off 2NA/λem plus m times the pattern frequency. It gives the Abbe-type resolution 1/k_c and the number of raw images, not the noise or the reconstruction quality.

  1. Gustafsson M. G. L. (2000). Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J. Microsc. 198, 82–87. Shifted passbands, k_c = 2NA/λem + k₀, three phases per angle. doi:10.1046/j.1365-2818.2000.00710.x
  2. Gustafsson M. G. L. (2005). Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution. PNAS 102, 13081–13086. Harmonics m·k₀ of saturated SIM. doi:10.1073/pnas.0406877102
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