Dynamic structured illumination microscopy: focused imaging and optical sectioning for moving objects

Leo G. Krzewina; Myung K. Kim

doi:10.1117/12.645411

23 February 2006 Dynamic structured illumination microscopy: focused imaging and optical sectioning for moving objects

Leo G. Krzewina, Myung K. Kim

Proceedings Volume 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII; 60900I (2006) https://doi.org/10.1117/12.645411
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

Structured illumination microscopy (SIM) is a valuable tool for three-dimensional microscopy and has numerous applications in bioscience. Its success has been limited to static objects, though, as three sequential image acquisitions are required per final processed, focused image. To overcome this problem we have developed a multicolored grid which when used in tandem with a color camera is capable of performing SIM with just a single exposure. Images and movies demonstrating optical sectioning of three-dimensional objects are presented, and results of applying color SIM for wide-field focused imaging are compared to those of SIM. From computer modeling and analytical calculations a theoretical estimate of the maximum observable object velocity in both the lateral and axial directions is available, implying that the new method will be capable of imaging a variety of live objects. Sample images of the technique applied to lens paper and a pigeon feather are included to show both advantages and disadvantages of CSIM.

Citation Download Citation

Leo G. Krzewina and Myung K. Kim "Dynamic structured illumination microscopy: focused imaging and optical sectioning for moving objects", Proc. SPIE 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII, 60900I (23 February 2006); https://doi.org/10.1117/12.645411

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