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Non-Scanning Holographic Fluorescence Micros
Non-Scanning Holographic Fluorescence Micros
Justine Dupere, Yamil Nieves, David C. Clark, Dept. of Physics, Univ. of South Florida
Initial Holograms
Final Holograms
Differential hologram focused at
stationary plane
Stationary plane (zh= -370mm)
Abstract:
Background:
(a)
(b)
(a)
Initial plane (zh= -1mm)
(c)
(b)
FluoSpheres Polystyrene Microspheres were
used from LifeTechnologies. Theyre 1 m and
excite/emit at 540/560nm. The beads were
mixed with a polyacrylamide gel to not only
prevent the beads from moving and clumping
together but to create a 3D matrix. (a), (b), (c)
and (c) show particles coming into focus at
different planes.
(d)
(e)
Experiments/Results:
Differential Holography:
Apparatus:
(f)
(g)
The fluorescent beads were shifted [-30.0x, -50.8y, 25.4z] m. The final
x,y position is indicated by the red circles in the dynamic plane-propagated
holograms. The bottom layer is a gel matrix and the top is a translating
layer, both with fluorescent beads. (a), (b), and (c) are the initial, final, and
differential holograms, respectively, at the stationary plane. (d) is the initial
position of the beads and (e) is the final position at the translated planes.
(f) and (g) are the differential holograms. The initial and final holograms
were subtracted, cancelling out the stationary information leaving only the
initial and final positions of the beads. So, the difference hologram is
focused to the stationary plane (c), the initial plane (f), and the final plane
(g).
Future Work:
(a)
(b)
(c)