A.O.T.F Laser control

• Thermally controlled Tellerium Dioxide (TeO2) crystal

Selected Wavelength

Kr/Ar Laser Light

Transducer

Non DefractedWavelengths

Acoustic Termination

BeamDump

Acousto-Optic-Tunable-Filter Principle

Specifications

• Polarisation preserving >70:1 extinction

• > 85% transmision efficiency

• blocking > OD 4.0

• Noise = < 5.0% peak RMS (measured in reflection imaging mode as per std. commissioning spec.)

• switching frequency - how quick can you click a mouse button!

AOTF DELIVERS High quality simultaneous multi label imaging

•A comparison of classic Neutral Density (ND) attenuation versus AOTF laser control

Spectral separation considerations

AOTF allows more control of the laser intensities to allowbetter assessment of dye co-localisation

FITC channel (522/32) using ND filter

CY3 channel (HQ598/40) using ND filter

CY5 channel (680/32) using ND filter

The FITC channel is bleeding into the CY3 channel in this set-up

24 Bit merge of FITC/CY3/CY5 using ND filter

Cross talk between red and green channels

Fluorogram of FITC & CY3 channels showing FITC bleed through (cross talk)

FITC channel (522/32) using AOTF control

CY3 channel (HQ598/40) using AOTF control

CY5 channel (680/32) using AOTF control

24 Bit merge of FITC/CY3/CY5 using AOTF control

Fluorogram of FITC & CY3 channels with AOTF control - displaying little co-localisation

The Comparison Again

AOTF imaging ND filter imaging

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