Structured illumination, often referred to as a "poor man's confocal microscope" is emerging as a powerful technique for optical sectioning in widefield microscopy at high resolution. Although current implementations are limited in speed and multi-channel acquisition by the requirement of capturing multiple images, new technological innovations are occurring rapidly in this field. The references listed in this section point to original research reports and review articles that should provide the starting point for a thorough understanding of structured illumination.
Neil, M. A. A., Juskaitis, R., and Wilson, T.
Method of obtaining optical sectioning by using structured light in a conventional microscope. Optics Letters 22: 1905-1907 (1997). The original research report on structured illumination that describes projecting a grid pattern onto the specimen at the focal plane. Included is a thorough mathematical description of the technique, microscope configuration, axial response, and images of a pollen grain.
Neil, M. A. A., Juskaitis, R., and Wilson, T.
Real time 3D fluorescence microscopy by two beam interference illumination. Optics Communications 153: 1-4 (1998). The demonstration of optical sectioning using structured illumination coupled to a standard widefield microscope. The investigators utilized two interfering beams on the specimen to create a single spatial frequency fringe pattern. The resulting images taken at three spatial positions of the fringe patterns were processed to produce optical sections.
Schaefer, L. H., Schuster, D., and Schaffer, J.
Structured illumination microscopy: artefact analysis and reduction utilizing a parameter optimization approach. Journal of Microscopy 216: 165-174 (2004). The authors describe practical applications of structured illumination microscopy and review potential artifacts. Among the most serious are residual stripe patterns originating from the illumination grating. Suggestions for correction of this artifact are provided.
Gustafsson, M. G. L.
Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. Journal of Microscopy 198: 82-87 (2000). An elegant discussion of structured illumination coupled to demonstrations of the high resolution capabilities exhibited by the technique. The author provides an impressive display of enhanced resolution using stress fibers forming the actin cytoskeleton at the edge of a cultured cell.
Gustafsson, M. G. L.
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution. Proceedings of the National Academy of Sciences, USA 102: 13081-13086 (2005). Experimental demonstration of the high resolution afforded by structured illumination coupled to laser illumination. The concept is thoroughly described and experimentally tested using fluorescent beads.
Neil, M. A. A., Wilson, T., and Juskaitis, R.
A light efficient optically sectioning microscope. Journal of Microscopy 189: 114-117 (1998). The authors describe the theoretical and practical aspects of configuring a standard widefield microscope for reflected light structured illumination. Included is a mathematical description, microscope configuration details, and example images of a semiconductor wafer surface.
Fedosseev, R., Belyaev, Y., Frohn, J., and Stemmer, A.
Structured light illumination for extended resolution in fluorescence microscopy. Optics and Lasers in Engineering 43: 403-414 (2005). A discussion of how structured illumination can extend optical resolution due to spatial frequencies beyond the classical cut-off frequency being brought into the microscope passband by frequency mixing, termed harmonic excitation light microscopy (HELM).
Gustafsson, M. G. L., Shao, L., Carlton, P. M., Wang, C. J. R., Golubovskaya, I. N., Cande, W. Z., Agard, D. A., and Sedat, J. W.
Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination. Biophysical Journal 94: 4957-4970 (2008). A brilliant description of how structured illumination can be applied in three dimensions to double the axial as well as lateral resolution in order to provide true optical sectioning. The authors give a complete theoretical description with impressive demonstrations using fluorescent beads and stained fixed specimens.
Schermelleh, L., Carlton, P. M., Haase, S., Shao, L., Winoto, L., Kner, P., Burke, B., Cardoso, M. C., Agard, D. A., Gustafsson, M. G. L., Leonhardt, H., and Sedat, J. W.
Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy. Science 320: 1332-1336 (2008). The authors describe the application of three-dimensional structured illumination microscopy (3D-SIM) to the observation of chromatin, nuclear lamina, and the nuclear pore complex using multiple fluorophores in fixed cells.
Poher, v., Zhang, H. X., Kennedy, G. T., Griffin, C., Oddos, S., Gu, E., Elson, D. S., Girkin, M., French, P. M. W., Dawson, M. D., and Neil, M. A.
Optical sectioning microscopes with no moving parts using a micro-stripe array light emitting diode. Optics Express 15: 11196-11206 (2007). A report describing structured illumination microscopy using a controlled LED array in place of a grid in the objective rear focal plane. The article describes the light source and microscope configuration, as well as a discussion of structured illumination microscopy. Included are specimen images and a mathematical description of the technology.