3-DIMENSIONAL HOLOGRAPHIC FLUORESCENCE MICROSCOPY

Most commonly used methods for three-dimensional (3D) fluorescence mic roscopy make use of sectioning techniques that require that the object be physically scanned in a series of two-dimensional (2D) sections al ong the z axis. The main drawback in these approaches is the need for these sequential 2D scans. An alternative approach to fluorescence ima ging in three dimensions has been developed that is based on optical s canning holography. This novel approach requires only a 2D scan to rec ord 3D information. Holograms of 15-mu m fluorescent latex beads with longitinal separation of similar to 2 mm have been recorded and recons tructed. To our knowledge, this is the first time holograms of fluores cent specimens have been recorded by an optical holographic technique. (C) 1997 Optical Society of America.