Full-field optical sectioning and three-dimensional localization of fluorescent particles using focal plane modulation.
We present a technique for imaging fluorescent particles based on the axial modulation of the objective's focal plane position. This technique provides full-field optical sectioning and can be used to localize the fluorophores in three dimensions. We describe the technique and apply it to image 200 nm diameter fluorescent beads immobilized in a gel. We show that full-field optical sectioning is obtained and that the beads are localized with a precision of 10 nm in the transverse plane and 14 nm in the axial direction.
Chasles
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F
Laboratoire d'Optique Physique, ESPCI, Paris, France. frederic.chasles@espci.fr
Dubertret
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B
Boccara
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AC