TY - JOUR
T1 - Widefield fluorescence microscopy with extended resolution
AU - Stemmer, Andreas
AU - Beck, Markus
AU - Fiolka, Reto
PY - 2008/11
Y1 - 2008/11
N2 - Widefield fluorescence microscopy is seeing dramatic improvements in resolution, reaching today 100 nm in all three dimensions. This gain in resolution is achieved by dispensing with uniform Köhler illumination. Instead, non-uniform excitation light patterns with sinusoidal intensity variations in one, two, or three dimensions are applied combined with powerful image reconstruction techniques. Taking advantage of non-linear fluorophore response to the excitation field, the resolution can be further improved down to several 10 nm. In this review article, we describe the image formation in the microscope and computational reconstruction of the high-resolution dataset when exciting the specimen with a harmonic light pattern conveniently generated by interfering laser beams forming standing waves. We will also discuss extensions to total internal reflection microscopy, non-linear microscopy, and three-dimensional imaging.
AB - Widefield fluorescence microscopy is seeing dramatic improvements in resolution, reaching today 100 nm in all three dimensions. This gain in resolution is achieved by dispensing with uniform Köhler illumination. Instead, non-uniform excitation light patterns with sinusoidal intensity variations in one, two, or three dimensions are applied combined with powerful image reconstruction techniques. Taking advantage of non-linear fluorophore response to the excitation field, the resolution can be further improved down to several 10 nm. In this review article, we describe the image formation in the microscope and computational reconstruction of the high-resolution dataset when exciting the specimen with a harmonic light pattern conveniently generated by interfering laser beams forming standing waves. We will also discuss extensions to total internal reflection microscopy, non-linear microscopy, and three-dimensional imaging.
KW - Extended resolution
KW - Structured illumination
KW - Widefield fluorescence microscopy
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U2 - 10.1007/s00418-008-0506-8
DO - 10.1007/s00418-008-0506-8
M3 - Review article
C2 - 18810482
AN - SCOPUS:54049117106
VL - 130
SP - 807
EP - 817
JO - Zeitschrift für Zellforschung und Mikroskopische Anatomie. Abteilung Histochemie
JF - Zeitschrift für Zellforschung und Mikroskopische Anatomie. Abteilung Histochemie
SN - 0948-6143
IS - 5
ER -