Structured Illumination Light Sheet Microscopy information
Structured-illumination light sheet microscopy
Structured illumination light sheet microscopy (SI-LSM) is an optical imaging technique used for achieving volumetric imaging with high temporal and spatial resolution in all three dimensions. It combines the ability of light sheet microscopy to maintain spatial resolution throughout relatively thick samples with the higher axial and spatial resolution characteristic of structured illumination microscopy. SI-LSM can achieve lateral resolution below 100 nm[1] in biological samples hundreds of micrometers thick.[2]
SI-LSM is most often used for fluorescent imaging of living biological samples, such as cell cultures. It is particularly useful for longitudinal studies, where high-rate imaging must be performed over long periods of time without damaging the sample.[3][4] The two methods most used for fluorescent imaging of 3D samples – confocal microscopy and widefield microscopy – both have significant drawbacks for this type of application. In widefield microscopy, both in-focus light from the plane of interest as well as out-of-focus light from the rest of the sample is acquired together, creating the “missing cone problem” which makes high resolution imaging difficult.[5] Although confocal microscopy largely solves this problem by using a pinhole to block unfocused light, this technique also inevitably blocks useful signal, which is particularly detrimental in fluorescent imaging when the signal is already very weak.[6] In addition, both widefield and confocal microscopy illuminate the entirety of the sample throughout imaging, which leads to problems with photobleaching and phototoxicity in some samples.[3][4] While light-field microscopy alone can address most of these issues, its achieved resolution is still fundamentally limited by the diffraction of light and it is unable to achieve super-resolution.[3][5]
SI-LSM works by using a patterned rather than uniform light sheet to illuminate a single plane of a volume being imaged. In this way, it maintains the many benefits of light-sheet microscopy while achieving the high resolution of structured illumination microscopy.
^Chang, Bo-Jui; Perez Meza, Victor Didier; Stelzer, Ernst H. K. (9 May 2017). "csiLSFM combines light-sheet fluorescence microscopy and coherent structured illumination for a lateral resolution below 100 nm". Proceedings of the National Academy of Sciences. 114 (19): 4869–4874. doi:10.1073/pnas.1609278114. PMC 5441750. PMID 28438995.
^Keller, Philipp J; Schmidt, Annette D; Santella, Anthony; Khairy, Khaled; Bao, Zhirong; Wittbrodt, Joachim; Stelzer, Ernst H K (August 2010). "Fast, high-contrast imaging of animal development with scanned light sheet–based structured-illumination microscopy". Nature Methods. 7 (8): 637–642. doi:10.1038/nmeth.1476. PMC 4418465. PMID 20601950.
^ abcGao, Liang; Shao, Lin; Higgins, Christopher D.; Poulton, John S.; Peifer, Mark; Davidson, Michael W.; Wu, Xufeng; Goldstein, Bob; Betzig, Eric (December 2012). "Noninvasive Imaging beyond the Diffraction Limit of 3D Dynamics in Thickly Fluorescent Specimens". Cell. 151 (6): 1370–1385. doi:10.1016/j.cell.2012.10.008. PMC 3615549. PMID 23217717.
^ abChen, Bi-Chang; Legant, Wesley R.; Wang, Kai; Shao, Lin; Milkie, Daniel E.; Davidson, Michael W.; Janetopoulos, Chris; Wu, Xufeng S.; Hammer, John A.; Liu, Zhe; English, Brian P.; Mimori-Kiyosue, Yuko; Romero, Daniel P.; Ritter, Alex T.; Lippincott-Schwartz, Jennifer; Fritz-Laylin, Lillian; Mullins, R. Dyche; Mitchell, Diana M.; Bembenek, Joshua N.; Reymann, Anne-Cecile; Böhme, Ralph; Grill, Stephan W.; Wang, Jennifer T.; Seydoux, Geraldine; Tulu, U. Serdar; Kiehart, Daniel P.; Betzig, Eric (24 October 2014). "Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution". Science. 346 (6208): 1257998. doi:10.1126/science.1257998. PMC 4336192. PMID 25342811.
^ abFrohn, J. T.; Knapp, H. F.; Stemmer, A. (1 June 2001). "Three-dimensional resolution enhancement in fluorescence microscopy by harmonic excitation". Optics Letters. 26 (11): 828–830. doi:10.1364/ol.26.000828. PMID 18040464.
^Gustafsson, M. G. L. (May 2000). "Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy". Journal of Microscopy. 198 (2): 82–87. doi:10.1046/j.1365-2818.2000.00710.x. PMID 10810003. S2CID 9257781.
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