Tag Archives: nanoscopy

Passive nanorheology measurements

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What do marshmallows, jelly (or Jell-O), cream cheese and Chinese soup dumplings have in common?  They are often made with gelatin.  Gelatin is derived from the skin and bones of cattle and pigs through the partial hydrolysis of collagen.  Gelatin is a physical hydrogel meaning that it consists of a three-dimensional network of polymer molecules in which a large amount of water is absorbed, as much as 90% in gelatin.  These polymer molecules are cross-linked by hydrogen bonds, hydrophobic interactions and chain entanglements.  External stimuli, such as temperature, can change the level of cross-linking causing the material to transition between its solid, liquid and gel states.  This is why jelly sets in the fridge and melts when it’s heated up – the cross-links holding the molecules together break down.  This type of responsive behaviour allows the properties of hydrogels to be controlled at the micro and sub-micron scale for a host of applications including tissue engineering, drug delivery, water treatment, wearable technologies, and supercapacitors.  However, the design and manufacture of soft hydrogels can be challenging due to the lack of technology for measuring the local properties.  Current quantitative techniques for measuring the properties of hydrogels usually focus on bulk properties and provide little data about local variations or real-time responses to external stimuli.  My colleagues and I have used gold nanoparticles as probes in hydrogels to map the properties at the microscale of thermosensitive hydrogels undergoing real-time transition from the solid to gel phases [see ‘Passive nanorheological tool to characterise hydrogels’].  This is an extension, or perhaps more accurately an application, of our earlier work on tracking nanoparticles through the vitreous humour of the eye [see ‘Nanoparticle motion-through heterogeneous hydrogels’ on November 6th, 2024].  The novel technique, which yields passive nanorheological measurements, allows us to evaluate local viscosity, identify time-varying heterogeniety and monitor dynamic phase transitions at the micro through to nano scale.  The significant challenges of other techniques, such as weak signals due to high water content and the dynamism of hydrogels, are overcome with a fast, inexpensive and user-friendly technology.  Although, even with these advantages, you are unlikely to use it when you are making jelly or roasting marshmallows over the campfire; however, it is really useful for understanding the transport of drugs through biological hydrogels or designing manufacturing processes for artificial tissue.

Reference

Moira Lorenzo Lopez, Victoria R. Kearns, Eann A. Patterson & Judith M. Curran, Passive nanorheological tool to characterise hydrogels, Nanoscale, 2025,17, 15338-15347.

Image: Figure 5 from the above reference showing a hydrogel transitioning to a gel phase as result of an increase in temperature with 100 nm diameter gold nanoparticles with some particles (yellow arrows) at the interface between phases.  The image was taken in an inverted optical microscope set up for tracking the nanoparticles.

Label-free real-time tracking of individual bacterium

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Images from the optical microscope showing the tracks of bacteria interacting with a surfaceAntimicrobial resistant (AMR) infections are already the third leading cause of death in the USA and are predicted to kill 50 million people per year by 2050.  It is the next pandemic starting already.  We have been using our capability to track nanoparticles in an optical microscope [see ‘Slow moving nanoparticles‘ on December 13th, 2017 and ‘Nano biomechanical engineering of agent delivery to cells‘ on December 15th, 2021] to track individual bacterium as they interact with surfaces to form biofilms.  Bacterial biofilms are complex colonies of bacteria that are highly resistant to antimicrobial agents and can cause life-threatening infections.  We have used our label-free, real-time tracking capabilities to explore the dynamics and adhesion of bacteria to surfaces and found that viable bacteria adhered to the surface but continue to move with rotary or sliding motions depending on the mechanics of their attachment to the surface.  Bacteria that were killed by contact with the surface did not move once they were attached to the surface.  The image shows examples of these motions from our paper published last month.  Our ability to detect these differences in the dynamics of bacteria will allow us to detect the onset of the formation of biofilms and to quantify the efficacy of antimicrobial surfaces and coatings.

Image: Figure 4 – Tracks (yellow lines) of the sections (purple circles) of four E. coli bacteria experiencing: (a) random diffusion above the surface; (b) rotary attachment; (c) lateral attachment; (d) static attachment. The dynamics of the four bacteria was monitored for approximately 20 s. The length of the scale bars is 5 μm. From Scientific Reports, 12:18146, 2022.

Source:

Giorgi F, Curran JM & Patterson EA, Real-time monitoring of the dynamics and interactions of bacteria and the early-stage formation of biofilms, Scientific Reports, 12:18146, 2022.

Size matters

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Most of us have a sub-conscious understanding of the forces that control the interaction of objects in the size scale in which we exist, i.e. from millimetres through to metres.  In this size scale gravitational and inertial forces dominate the interactions of bodies.  However, at the size scale that we cannot see, even when we use an optical microscope, the forces that the dominate the behaviour of objects interacting with one another are different.  There was a hint of this change in behaviour observed in our studies of the diffusion of nanoparticles [see ‘Slow moving nanoparticles‘ on December 13th, 2017], when we found that the movement of nanoparticles less than 100 nanometres in diameter was independent of their size.  Last month we published another article in one of the Nature journals, Scientific Reports, on ‘The influence of inter-particle forces on diffusion at the nanoscale‘, in which we have demonstrated by experiment that Van der Waals forces and electrostatic forces are the dominant forces at the nanoscale.  These forces control the diffusion of nanoparticles as well as surface adhesion, friction and colloid stability.  This finding is significant because the ionic strength of the medium in which the particles are moving will influence the strength of these forces and hence the behaviour of the nanopartices.  Since biological fluids contain ions, this will be important in understanding and predicting the behaviour of nanoparticles in biological applications where they might be used for drug delivery, or have a toxicological impact, depending on their composition.

Van der Waals forces are weak attractive forces between uncharged molecules that are distance dependent.  They are named after a Dutch physicist, Johannes Diderik van der Waals (1837-1923).  Electrostatic forces occur between charged particles or molecules and are usually repulsive with the result that van der Waals and electrostatic forces can balance each other, or not depending on the circumstances.

Sources:

Giorgi F, Coglitore D, Curran JM, Gilliland D, Macko P, Whelan M, Worth A & Patterson EA, The influence of inter-particle forces on diffusion at the nanoscale, Scientific Reports, 9:12689, 2019.

Coglitore D, Edwardson SP, Macko P, Patterson EA, Whelan MP, Transition from fractional to classical Stokes-Einstein behaviour in simple fluids, Royal Society Open Science, 4:170507, 2017. doi: 10.1098/rsos.170507.

Patterson EA & Whelan MP, Tracking nanoparticles in an optical microscope using caustics. Nanotechnology, 19 (10): 105502, 2009.

Image: from Giorgi et al 2019, figure 1 showing a photograph of a caustic (top) generated by a 50 nm gold nanoparticle in water taken with the optical microscope adjusted for Kohler illumination and closing the condenser field aperture to its minimum following method of Patterson and Whelan with its 2d random walk over a period of 3 seconds superimposed and a plot of the same walk (bottom).

Assessing nanoparticle populations in historic nuclear waste

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Together with colleagues at the JRC Ispra, my research group has shown that the motion of small nanoparticles at low concentrations is independent of their size, density and material [1], [see ‘Slow moving nanoparticles‘ on December 13th, 2017].  This means that commercially-available instruments for evaluating the size and number of nanoparticles in a solution will give erroneous results under certain conditions.  In a proposed PhD project, we are planning to extend our work to develop an instrument with capability to automatically identify and size nanoparticles, in the range from 1 to 150 nanometres, using the three-dimensional optical signature, or caustic, which particles generate in an optical microscope, that can be several orders of magnitude larger than the particle [2],  [see ‘Toxic nanoparticles?‘ on November 13th, 2013].  The motivation for the work is the need to characterise particles present in solution in legacy ponds at Sellafield.  Legacy ponds at the Sellafield site have been used to store historic radioactive waste for decades and progress is being made in reducing the risks associated with these facilities [3].  Over time, there has been a deterioration in the condition of the ponds and their contents that has resulted in particles being present in solution in the ponds.  It is important to characterise these particles in order to facilitate reductions in the risks associated with the ponds.  We plan to use our existing facilities at the University of Liverpool to develop a novel instrument using simple solutions probably with gold nanoparticles and then to progress to non-radioactive simulants of the pond solutions.  The long-term goal will be to transition the technology to the Sellafield site perhaps with an intermediate step involving a demonstration of  the technology on pond solutions using the facilities of the National Nuclear Laboratory.

The PhD project is fully-funded for UK and EU citizens as part of a Centre for Doctoral Training and will involve a year of specialist training followed by three years of research.  For more information following this link.

References:

[1] Coglitore, D., Edwardson, S.P., Macko, P., Patterson, E.A., & Whelan, M.P., Transition from fractional to classical Stokes-Einstein behaviour in simple fluids, Royal Society Open Science, 4:170507, 2017.

[2] Patterson, E.A., Whelan, P., 2008, ‘Optical signatures of small nanoparticles in a conventional microscope’ Small, 4(10):1703-1706.

[3] Comptroller and Auditor General, The Nuclear Decommissioning Authority: progress with reducing risk at Sellafield, National Audit Office, HC 1126, Session 2017-19, 20 June 2018.