TY - JOUR
T1 - Clearer view for TIRF and oblique illumination microscopy
AU - Fiolka, Reto
N1 - Publisher Copyright:
© 2016 Optical Society of America.
PY - 2016/12/26
Y1 - 2016/12/26
N2 - In Total Internal Reflection Fluorescence (TIRF) microscopy, the sample is illuminated with an evanescent field that yields a thin optical section. However, its widefield detection has no rejection mechanism against out-of-focus blur from scattered light that can compromise TIRF images. Here I demonstrate that via structured illumination, out-of-focus blur can be effectively suppressed in TIRF microscopy, yielding strikingly clearer images. The same mechanism can also be applied to oblique illumination schemes that extend the reach of TIRF microscopy beyond the basal surface of the cell. The two imaging modes are used to image a biosensor, clathrin coated vesicles and the actin cytoskeleton in different cell types with improved contrast.
AB - In Total Internal Reflection Fluorescence (TIRF) microscopy, the sample is illuminated with an evanescent field that yields a thin optical section. However, its widefield detection has no rejection mechanism against out-of-focus blur from scattered light that can compromise TIRF images. Here I demonstrate that via structured illumination, out-of-focus blur can be effectively suppressed in TIRF microscopy, yielding strikingly clearer images. The same mechanism can also be applied to oblique illumination schemes that extend the reach of TIRF microscopy beyond the basal surface of the cell. The two imaging modes are used to image a biosensor, clathrin coated vesicles and the actin cytoskeleton in different cell types with improved contrast.
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U2 - 10.1364/OE.24.029556
DO - 10.1364/OE.24.029556
M3 - Article
C2 - 28059342
AN - SCOPUS:85009350754
SN - 1094-4087
VL - 24
SP - 29556
EP - 29567
JO - Optics Express
JF - Optics Express
IS - 26
ER -