Abstract

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 200nm 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 10nm in the transverse plane and 14nm in the axial direction.

© 2006 Optical Society of America

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