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Bató Lilia

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Bató Lilia
An obstacle-free microfluidic system for monitoring protein diffusion

Aug 30 - szerda

11:00 – 11:15

Bioszenzorika és bio-nanotechnológia

E26

An obstacle-free microfluidic system for monitoring protein diffusion

Lilia Bató1,2, Péter Fürjes1

1 Microsystems Lab., Inst. of Technical Physics and Materials Science, Centre for Energy Research, ELKH, Budapest, Hungary

2 Óbuda University Doctoral School on Materials Sciences and Technologies, Budapest, Hungary

Determining characteristic chemical and physical properties of proteins are essential for predicting their behaviour and interactions in Lab-on-Chip and Organ-on-Chip systems. Diffusion coefficients are widely used to estimate molecule size, hydration state or aggregation [1, 2]. Constructed diffusion is crucial to develop adequate molecular concentration distribution in these applications. Our goal is to develop a free-diffusion based obstacle-free microfluidic device to achieve fast diffusion coefficient measurements.

A specific microfluidic system was created – inspired by Taylor et al. [3] – containing two main channels connected by multiple perpendicular capillaries to ensure stable and stationary molecular concentration at the edge of the monitoring channel. The device was fabricated by soft lithography in polydimethylsiloxane and was bonded to a glass slide. The main channels were filled with test puffer and adequate fluorescent protein solutions, respectively. The symmetry of the layout minimizes the pressure differences between the channels.

Diffusion coefficient of various, fluorescently labelled proteins (bovine serum albumin, immunoglobulin G, rhodamine B) were determined using time dependent fluorescent microscopy. To acquire the evolving intensity profiles along the channels 30 s time-lapse images were taken. Python evaluation code was developed to fit model complementary error functions to the experimental profiles and calculate the diffusion coefficients. The defined diffusion coefficients for the certain proteins were in accordance with the values found in literature and estimated by the Stokes-Einstein equation.

Easy-to-use free-diffusion based microfluidic system were developed and proved to be applicable for fast determination the diffusion coefficients of characteristic proteins, using time-lapse fluorescent imaging and efficient evaluation algorithm.

Acknowledgement

The work was supported by the National Research, Development and Innovation Fund (NKFIA) via INBIOM TKP2021-EGA-04 grant.

References

  • [1] P. Arosio et al, ACS Nano, 10: 333-341, 2015
  • [2] M. R. G. Kopp and P. Arosio, J. Pharm. Sci, 107: 1228-1236, 2018
  • [3] Taylor et al. Nat. Methods, 8:599-605, 2005