Skip to main content
SpringerLink
Log in

High-speed velocimetry in microfluidic protein mixers using confocal fluorescence decay microscopy

  • Research Article
  • Published:
Experiments in Fluids Aims and scope Submit manuscript

Abstract

A method to measure fluid speeds on the order of 10,000 mm/s in microchannels is presented. A microfluidic protein mixer is manufactured with a 170 µm microscope coverslip bottom that interfaces with a confocal florescence microscope using a water-immersed Olympus UPLSAPO 60XW objective to create a diffraction-limited confocal volume. A diode laser with a repetition rate of 1 MHz is used to study Poiseuille flows at average speeds of 5000, 6000 and 7000 mm/s by exciting tris(2,2′-bipyridine)ruthenium(II) hexafluorophosphate solution at 1 μmol/L concentration flowing through the micro-mixer in the confocal volume. Decays collected using a time-correlated single photon counting card at each grid point are characterized by the first moment of the decay and curve fitted with the theoretical Poiseuille flow solutions. It was found that curve fitting with higher average speeds results in lower errors. A fluorescence correlation study was then carried out at different depths in the micro-mixer to understand the raw data profiles observed using the diode laser. A mixing study was then carried out using a Ti-Sapphire laser with a repetition rate of 3.8 MHz. A Poiseuille flow at 7000 mm/s was measured using the Ti-Sapphire laser and then curve fitted to the theoretical Poiseuille flow solution. The curve fit was then applied to the complicated flow region to determine speed. Results of the experimental mixing study are also compared to direct numerical simulation results.

Graphical abstract

An experimental technique to measure speeds in the complicated mixing region of a protein mixer is developed. Results show that when flow is bounded asymmetrically between a rough wall and smooth wall, the flow shows affinity for the smooth wall during the mixing process. The Reynolds number for this is flow is 245 indicating flow in the transitional turbulence regime.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14
Fig. 15
Fig. 16
Fig. 17
Fig. 18

Similar content being viewed by others

References

  • Acton FS (1959) Analysis of straight-line data. Wiley, New York

    MATH  Google Scholar 

  • Bostock L, Chandler S, Rourke C (1982) Further pure mathematics. Nelson Thornes, Cheltenham, p 410

    Google Scholar 

  • Buschmann V, Krämer B, Koberling F, Macdonald R, Rättinger S (2009) Quantitative FCS: determination of the confocal volume by FCS and bead scanning with the microtime 200. Application Note PicoQuant GmbH, Berlin

    Google Scholar 

  • Courant R, Friedrichs K, Lewy H (1967) On the partial difference equations of mathematical physics. IBM J Res Dev 11:215–234

    Article  MathSciNet  Google Scholar 

  • Cox G, Sheppard CJ (2004) Practical limits of resolution in confocal and non-linear microscopy. Microsc Res Tech 63:18–22

    Article  Google Scholar 

  • Gobby D, Angeli P, Gavriilidis A (2001) Mixing characteristics of T-type microfluidic mixers. J Micromech Microeng 11:126

    Article  Google Scholar 

  • Inguva V, Kathuria SV, Bilsel O, Perot BJ (2018) Computer design of microfluidic mixers for protein/RNA folding studies. PLoS One 13:e0198534

    Article  Google Scholar 

  • Jiang X, Lai C (2016) Numerical techniques for direct and large-eddy simulations. CRC Press, Boca Raton

    MATH  Google Scholar 

  • Kathuria SV, Chan A, Graceffa R, Paul Nobrega R, Robert Matthews C, Irving TC, Perot B, Bilsel O (2013) Advances in turbulent mixing techniques to study microsecond protein folding reactions. Biopolymers 99:888–896

    Article  Google Scholar 

  • Kinoshita H, Kaneda S, Fujii T, Oshima M (2007) Three-dimensional measurement and visualization of internal flow of a moving droplet using confocal micro-PIV. Lab Chip 7:338–346

    Article  Google Scholar 

  • Ku HH (1966) Notes on the use of propagation of error formulas. J Res Natl Bur Stand 70:263–273

    Google Scholar 

  • Li H, Olsen MG (2006) Examination of large-scale structures in turbulent microchannel flow. Exp Fluids 40:733–743

    Article  Google Scholar 

  • Majumdar ZK, Sutin JD, Clegg RM (2005) Microfabricated continuous-flow, turbulent, microsecond mixer. Rev Sci Instrum 76:125103

    Article  Google Scholar 

  • Martell MB, Rothstein JP, Perot JB (2010) An analysis of superhydrophobic turbulent drag reduction mechanisms using direct numerical simulation. Phys Fluids 22:065102

    Article  Google Scholar 

  • Matsumoto S, Yane A, Nakashima S, Hashida M, Fujita M, Goto Y, Takahashi S (2007) A rapid flow mixer with 11-µs mixing time microfabricated by a pulsed-laser ablation technique: observation of a barrier-limited collapse in cytochrome c folding. J Am Chem Soc 129:3840–3841

    Article  Google Scholar 

  • Natrajan V, Christensen K (2009) Structural characteristics of transition to turbulence in microscale capillaries. Phys Fluids 21:034104

    Article  Google Scholar 

  • Nguyen N (2011) Micromixers: fundamentals, design and fabrication. Elsevier, Oxford, pp 28–36

    Google Scholar 

  • Park JS, Choi CK, Kihm KD (2004) Optically sliced micro-PIV using confocal laser scanning microscopy (CLSM). Exp Fluids 37:105–119

    Article  Google Scholar 

  • Perot JB (2011) Determination of the decay exponent in mechanically stirred isotropic turbulence. AIP Adv 1:022104

    Article  Google Scholar 

  • Pope SB (2001a) The scales of turbulent motion. In: Turbulent flows, pp 184–187

  • Pope SB (2001b) Wall flows. In: Turbulent flows, p 270

  • Schermelleh L, Heintzmann R, Leonhardt H (2010) A guide to super-resolution fluorescence microscopy. J Cell Biol 190:165–175. https://doi.org/10.1083/jcb.201002018

    Article  Google Scholar 

  • Shinohara K, Sugii Y, Aota A, Hibara A, Tokeshi M, Kitamori T, Okamoto K (2004) High-speed micro-PIV measurements of transient flow in microfluidic devices. Meas Sci Technol 15:1965

    Article  Google Scholar 

  • Tanaami T, Otsuki S, Tomosada N, Kosugi Y, Shimizu M, Ishida H (2002) High-speed 1-frame/ms scanning confocal microscope with a microlens and Nipkow disks. Appl Opt 41:4704–4708

    Article  Google Scholar 

  • Van Houten J, Watts RJ (1976) Temperature dependence of the photophysical and photochemical properties of the tris (2,2′-bipyridyl)ruthenium(II) ion in aqueous solution. J Am Chem Soc 98:4853–4858

    Article  Google Scholar 

  • Van Oort B, Amunts A, Borst JW, Van Hoek A, Nelson N, Van Amerongen H, Croce R (2008) Picosecond fluorescence of intact and dissolved PSI-LHCI crystals. Biophys J 95:5851–5861

    Article  Google Scholar 

  • Williams SJ, Park C, Wereley ST (2010) Advances and applications on microfluidic velocimetry techniques. Microfluid Nanofluidics 8:709–726

    Article  Google Scholar 

  • Zettner C, Yoda M (2003) Particle velocity field measurements in a near-wall flow using evanescent wave illumination. Exp Fluids 34:115–121

    Article  Google Scholar 

  • Zusi CJ, Perot JB (2013) Simulation and modeling of turbulence subjected to a period of uniform plane strain. Phys Fluids 25:110819

    Article  Google Scholar 

Download references

Acknowledgements

We would like to thank NSF for funding this work. The work is supported by NSF IDBR Award no. 1353942. We would like to thank Yvonne Chan at University of Massachusetts Medical School for providing assistance in the wet laboratory. We would like to thank the reviewers for their constructive feedback that significantly improved this paper. We acknowledge the Texas Advanced Computing Center (TACC) at The University of Texas at Austin for providing HPC, visualization and database resources that have contributed to the research results reported within this paper. http://www.tacc.utexas.edu.

Author information

Authors and Affiliations

  1. University of Massachusetts Amherst, Amherst, MA, 01003, USA

    Venkatesh Inguva, Jonathan P. Rothstein & Blair J. Perot

  2. University of Massachusetts Medical School, Worcester, MA, 01655, USA

    Osman Bilsel

Authors
  1. Venkatesh Inguva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jonathan P. Rothstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Osman Bilsel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Blair J. Perot
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Venkatesh Inguva.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (ZIP 276393 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Inguva, V., Rothstein, J.P., Bilsel, O. et al. High-speed velocimetry in microfluidic protein mixers using confocal fluorescence decay microscopy. Exp Fluids 59, 177 (2018). https://doi.org/10.1007/s00348-018-2630-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00348-018-2630-0

Search

Navigation