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Time-resolved fluorescence imaging of solvent interactions in microfluidic devices

Richard K. P. Benninger, Oliver Hofmann, James McGinty, Jose Requejo-Isidro, Ian Munro, Mark A. A. Neil, Andrew J. deMello, and Paul M. W. French

Open Access Open Access

Abstract

We present the application of wide-field time-resolved fluorescence imaging methods for the study of solvent interactions and mixing in microfluidic devices. Time-resolved imaging of fluorescence polarization anisotropy allows us to image the local viscosity of fluorescein in three dimensions in order to directly monitor solvent mixing within a microfluidic channel. This provides a viscosity image acquisition time of the order of minutes, and has been applied to a steady-state laminar flow configuration. To image dynamic fluid mixing in real-time, we demonstrate high-speed fluorescence lifetime imaging at 12.3 Hz applied to DASPI, which directly exhibits a solvent viscosity-dependant fluorescence lifetime. These two methods facilitate a high degree of quantification of microfluidic flow in 3-D and/or at high speed, providing a tool for studying fluid dynamics and for developing enhanced microfluidic assays.

©2005 Optical Society of America

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Supplementary Material (2)

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Media 2: AVI (3419 KB)     

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Figures (8)
Fig. 1.
Fig. 1. Schematic of rFLIM setup

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Fig. 2.
Fig. 2. Schematic of high speed FLIM setup

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Fig. 3.
Fig. 3. (a) Channel schematic, (b) image of correlation time at junction and (c) Sample data points with fit

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Fig. 4.
Fig. 4. (a) and (b) Images of correlation time at ~0.4cm downstream of the point of confluence at 10 and 1 μl/min flow rate respectively. (c) Comparison of viscosity profiles across channel for the different flow rates.

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Fig. 5.
Fig. 5. Rendered 3D viscosity profile of flow interface, viewed from above.

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Fig. 6.
Fig. 6. Comparison of (a) rotational correlation time images, (b) fluorescence lifetime images and (c) profiles across the channel.

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Fig. 7.
Fig. 7. (3.33MB) Movie of fluorescence lifetime at point of confluence, with color scale attached. Flows were applied through the two side inlets while the center inlet was unused.

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Fig. 8.
Fig. 8. (3.33MB) Movie of fluorescence lifetime at channel 5cm from mixing point as flow is decreased from 10 to 0 μl/min. Color scale as in fig. 7.

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Equations (7)

Equations on this page are rendered with MathJax. Learn more.

r ( t ) = ⌊ I ∥ ( t ) − G I ⊥ ( t ) ⌋ [ I ∥ ( t ) + 2 G I ⊥ ( t ) ]
G = ( I VV I VH I HV I HH ) 1 2
I ∥ = K a I x + K b I y + K c I z I ⊥ = K a I x + K b I z + K c I y
r ( t ) = r 0 exp ( − t θ )
θ = V η 3 kT
τ RLD = Δ t ln ( I 1 I 2 )
F ( t ) = I ∥ ( t ) + 2 G I ⊥ ( t )
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