# Microrheology

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**Microrheology**[1] is a technique used to measure the [rheological properties](/source/Rheology) of a medium, such as [microviscosity](/source/Microviscosity), via the measurement of the trajectory of a [flow tracer](/source/Flow_tracer) (a [micrometre](/source/Micrometre)-sized particle). It is a new way of doing [rheology](/source/Rheology), traditionally done using a [rheometer](/source/Rheometer). There are two types of microrheology: *passive microrheology* and *active microrheology*. Passive microrheology uses inherent [thermal energy](/source/Thermal_energy) to move the tracers, whereas active microrheology uses externally applied forces, such as from a [magnetic field](/source/Magnetic_field) or an [optical tweezer](/source/Optical_tweezer), to do so. Microrheology can be further differentiated into 1- and 2-particle methods.[2] [3]

## Passive microrheology

*Passive microrheology* uses the thermal energy (*kT*) to move the tracers, although recent evidence suggests that active random forces inside cells may instead move the tracers in a diffusive-like manner.[4] The trajectories of the tracers are measured optically either by microscopy, or alternatively by light scattering techniques. [Diffusing-wave spectroscopy](/source/Diffusing-wave_spectroscopy) (DWS) is a common choice that extends light scattering measurement techniques to account for multiple scattering events.[5] From the [mean squared displacement](/source/Mean_squared_displacement) with respect to time (noted MSD or <Δ*r*2> ), one can calculate the visco-elastic moduli *G*′(*ω*) and *G*″(*ω*) using the [generalized Stokes–Einstein relation](/source/Einstein_relation_(kinetic_theory)) (GSER). Here is a view of the trajectory of a particle of micrometer size.

		- Typical trajectory of a Brownian particle (simulation)

		- Two examples of MSD: one for a purely viscous fluid (free diffusion) and one for a viscolelastic fluid (trapped by elastic network)

		- Animation of a particle in a polymer-like network

In a standard passive microrheology test, the movement of dozens of tracers is tracked in a single video frame. The motivation is to average the movements of the tracers and calculate a robust MSD profile.

Observing the MSD for a wide range of integration time scales (or frequencies) gives information on the microstructure of the medium where are diffusing the tracers.

If the tracers are experiencing free diffusion in a purely viscous material, the MSD should grow linearly with sampling integration time:

⟨ Δ r 2 ⟩ = 4 D t {\displaystyle \langle \Delta r^{2}\rangle =4Dt} .

If the tracers are moving in a spring-like fashion within a purely elastic material, the MSD should have no time dependence:

⟨ Δ r 2 ⟩ = Const {\displaystyle \langle \Delta r^{2}\rangle ={\text{Const}}}

In most cases the tracers are presenting a sub-linear integration-time dependence, indicating the medium has intermediate viscoelastic properties. Of course, the slope changes in different time scales, as the nature of the response from the material is frequency dependent.

Microrheology is another way to do linear rheology. Since the force involved is very weak (order of 10−15 N), microrheology is guaranteed to be in the so-called linear region of the strain/stress relationship. It is also able to measure very small volumes (biological cell).

Given the complex viscoelastic modulus G ( ω ) = G ′ ( ω ) + i G ″ ( ω ) {\displaystyle G(\omega )=G'(\omega )+iG''(\omega )\,} with *G*′(*ω*) the elastic (conservative) part and *G*″(*ω*) the viscous (dissipative) part and *ω*=2*πf* the pulsation. The GSER is as follows:

- G ~ ( s ) = k B T π a s ⟨ Δ r ~ 2 ( s ) ⟩ {\displaystyle {\tilde {G}}(s)={\frac {k_{\mathrm {B} }T}{\pi as\langle \Delta {\tilde {r}}^{2}(s)\rangle }}}

with

- G ~ ( s ) {\displaystyle {\tilde {G}}(s)} : [Laplace](/source/Laplace) transform of *G*

- *k*B: [Boltzmann](/source/Boltzmann) constant

- *T*: temperature in kelvins

- *s*: the Laplace frequency

- *a*: the radius of the tracer

- ⟨ Δ r ~ 2 ( s ) ⟩ {\displaystyle \langle \Delta {\tilde {r}}^{2}(s)\rangle } : the Laplace transform of the mean squared displacement

A related method of passive microrheology involves the tracking positions of a particle at high frequency, often with a quadrant photodiode.[6] From the position, x ( t ) {\displaystyle x(t)} , the power spectrum, ⟨ x ω 2 ⟩ {\displaystyle \langle x_{\omega }^{2}\rangle } can be found, and then related to the real and imaginary parts of the response function, α ( ω ) {\displaystyle \alpha (\omega )} .[7] The response function leads directly to a calculation of the complex shear modulus, G ( ω ) {\displaystyle G(\omega )} via:

- G ( ω ) = 1 6 π a α ( ω ) {\displaystyle G(\omega )={\frac {1}{6\pi a\alpha (\omega )}}}

## Two Point Microrheology

Source:[8]

There could be many artifacts that change the values measured by the passive microrheology tests, resulting in a disagreement between microrheology and normal [rheology](/source/Rheology). These artifacts include tracer-matrix interactions, tracer-matrix size mismatch and more.

A different microrheological approach studies the [cross-correlation](/source/Cross-correlation) of two tracers in the same sample. In practice, instead of measuring the MSD ⟨ Δ r 2 ⟩ {\displaystyle \langle \Delta r^{2}\rangle } , movements of two distinct particles are measured - ⟨ Δ r 1 Δ r 2 ⟩ {\displaystyle \langle \Delta r_{1}\Delta r_{2}\rangle } . Calculating the G(ω) of the medium between the tracers follows:

G ~ ( s ) = k B T 2 π R s ⟨ Δ r ~ 1 ( s ) Δ r ~ 2 ( s ) ⟩ {\displaystyle {\tilde {G}}(s)={\frac {k_{\mathrm {B} }T}{2\pi Rs\langle \Delta {\tilde {r}}_{1}(s)\Delta {\tilde {r}}_{2}(s)\rangle }}}

Notice this equation does not depend on **a**, but instead in depends on **R** - the distance between the tracers (assuming R>>a).

Some studies has shown that this method is better in coming to agreement with standard rheological measurements (in the relevant frequencies and materials)

## Active microrheology

*Active microrheology* may use a [magnetic field](/source/Magnetic_field) ,[9][10][11][8][12] [13] [optical tweezers](/source/Optical_tweezers)[14] [15] [16] [17][18] or an [atomic force microscope](/source/Atomic_force_microscopy)[19] to apply a force on the tracer and then find the stress/strain relation.

The force applied is a sinusoidal force with amplitude A and frequency ω -

F = A sin ⁡ ( ω t ) {\displaystyle F=A\sin(\omega t)}

The response of the tracer is a factor of the matrix visco-elastic nature. If a matrix is totally elastic (a solid), the response to the acting force should be immediate and the tracers should be observed moving by-

X e = B sin ⁡ ( ω t ) {\displaystyle X_{e}=B\sin(\omega t)} .

with A / B = G ( ω ) {\displaystyle A/B=G(\omega )} .

On the other hand, if the matrix is totally viscous (a liquid), there should be a phase shift of 90 o {\displaystyle 90^{o}} between the strain and the stress -

X v = B sin ⁡ ( ω t + 90 o ) = B cos ⁡ ( ω t ) {\displaystyle X_{v}=B\sin(\omega t+90^{o})=B\cos(\omega t)}

in reality, as most materials are visco-elastic, the phase shift observed is 0 < φ < 90 {\displaystyle 0<\varphi <90} .

When φ>45 the matrix is considered mostly in its "viscous domain" and when φ<45 the matrix is considered mostly in its "elastic domain".

Given a measured response phase shift φ (sometimes noted as δ), this ratio applies:

G ″ G ′ = A B sin ⁡ ( φ ) A B cos ⁡ ( φ ) = tan ⁡ ( φ ) {\displaystyle {\frac {G''}{G'}}={\frac {{\frac {A}{B}}\sin(\varphi )}{{\frac {A}{B}}\cos(\varphi )}}=\tan(\varphi )}

Similar response phase analysis is used in regular [rheology](/source/Rheology) testing.

More recently, it has been developed into [Force spectrum microscopy](/source/Force_spectrum_microscopy) to measure contributions of random active motor proteins to diffusive motion in the [cytoskeleton](/source/Cytoskeleton).[4]

## References

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1. ^ [***a***](#cite_ref-Guo_4-0) [***b***](#cite_ref-Guo_4-1) Guo, Ming; et al. (2014). ["Probing the Stochastic, Motor-Driven Properties of the Cytoplasm Using Force Spectrum Microscopy"](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4183065). *Cell*. **158** (4): 822–832. [doi](/source/Doi_(identifier)):[10.1016/j.cell.2014.06.051](https://doi.org/10.1016%2Fj.cell.2014.06.051). [PMC](/source/PMC_(identifier)) [4183065](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4183065). [PMID](/source/PMID_(identifier)) [25126787](https://pubmed.ncbi.nlm.nih.gov/25126787).

1. **[^](#cite_ref-5)** Furst, Eric M.; Todd M. Squires (2017). *Microrheology* (First ed.). Oxford. [ISBN](/source/ISBN_(identifier)) [978-0-19-965520-5](https://en.wikipedia.org/wiki/Special:BookSources/978-0-19-965520-5). [OCLC](/source/OCLC_(identifier)) [990115841](https://search.worldcat.org/oclc/990115841).{{[cite book](https://en.wikipedia.org/wiki/Template:Cite_book)}}: CS1 maint: location missing publisher ([link](https://en.wikipedia.org/wiki/Category:CS1_maint:_location_missing_publisher))

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1. **[^](#cite_ref-18)** David Engström; Michael C.M. Varney; Martin Persson; Rahul P. Trivedi; et al. (2012). ["Unconventional structure-assisted optical manipulation of high-index nanowires in liquid crystals"](https://doi.org/10.1364%2FOE.20.007741). *[Optics Express](/source/Optics_Express)*. **20** (7): 7741–7748. [Bibcode](/source/Bibcode_(identifier)):[2012OExpr..20.7741E](https://ui.adsabs.harvard.edu/abs/2012OExpr..20.7741E). [doi](/source/Doi_(identifier)):[10.1364/OE.20.007741](https://doi.org/10.1364%2FOE.20.007741). [PMID](/source/PMID_(identifier)) [22453452](https://pubmed.ncbi.nlm.nih.gov/22453452). [S2CID](/source/S2CID_(identifier)) [13975852](https://api.semanticscholar.org/CorpusID:13975852).

1. **[^](#cite_ref-19)** Rigato, Annafrancesca; Miyagi, Atsushi; Scheuring, Simon; Rico, Felix (2017-05-01). ["High-frequency microrheology reveals cytoskeleton dynamics in living cells"](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5540170). *Nature Physics*. advance online publication (8): 771–775. [Bibcode](/source/Bibcode_(identifier)):[2017NatPh..13..771R](https://ui.adsabs.harvard.edu/abs/2017NatPh..13..771R). [doi](/source/Doi_(identifier)):[10.1038/nphys4104](https://doi.org/10.1038%2Fnphys4104). [ISSN](/source/ISSN_(identifier)) [1745-2481](https://search.worldcat.org/issn/1745-2481). [PMC](/source/PMC_(identifier)) [5540170](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5540170). [PMID](/source/PMID_(identifier)) [28781604](https://pubmed.ncbi.nlm.nih.gov/28781604).

## External links

- [Harvard Weitz Lab page](https://web.archive.org/web/20090530054237/http://www.seas.harvard.edu/weitzlab/research/micrheo.html)

- [Review of microrheology in optical tweezers](http://www.rsc.org/delivery/_ArticleLinking/DisplayHTMLArticleforfree.cfm?JournalCode=LC&Year=2009&ManuscriptID=b907992k&Iss=17)

- [Review on microrheology](https://web.archive.org/web/20031010104303/http://www.deas.harvard.edu/projects/weitzlab/papers/urheo_chapter.pdf)

- [Illustrated description of microrheology and a microrheology analysis, with movies](https://archive.today/20111010061145/http://www.formulaction.com/microrheology_viscoelasticity.html)

- [DWS Microrheology Overview](https://lsinstruments.ch/en/technology/diffusing-wave-spectroscopy-dws/microrheology/)

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Adapted from the Wikipedia article [Microrheology](https://en.wikipedia.org/wiki/Microrheology) by Wikipedia contributors ([contributor history](https://en.wikipedia.org/wiki/Microrheology?action=history)). Available under [Creative Commons Attribution-ShareAlike 4.0 International](https://creativecommons.org/licenses/by-sa/4.0/). Changes may have been made.