Tag Archives: caustics

Fancy a pint of science?

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In September I am planning to initiate a new research project on the interaction of bacteria with cellular and hard surfaces.  It is in collaboration with Jude Curran and is co-funded by Unilever and the Biotechnology and Biological Sciences Research Council.  We have already used the optical method of caustics in a microscope to track and characterise the movement of synthetic nanoparticles as small as 3 nm in an array of biologically-relevant solutions [see ‘Nano biomechanical engineering of agent delivery to cells’ on December 15th, 2021].  We have also used the same technique to characterise and quantify the motion and growth of bacteria in solutions.  Now, we plan to use caustic signatures as a label-free tracking technology for pre-clinical testing of antimicrobial solutions and coatings.  We plan to start by considering binding and removal of viral particles and bacterial spores from hard and soft laundry surfaces using various bacterial species, including Staph aureus which is responsible for laundry malodour; before progressing to the interaction of bacteria with human oral and skin cell cultures.  We are in the process of recruiting a suitable PhD student so if you are interested or know someone who might be suitable then get in touch.  If you want to learn more about our tracking technology and fancy a pint of science, then join us in Liverpool in May for part of the world’s largest festival of public science.  I will be talking about ‘Revealing the invisible: real-time motion of virus particles’  on May 10th at 7.30pm on Leaf of Bold Street.

Liverpool Pint of Science programme

UK Pint of Science programme

 

Nano biomechanical engineering of agent delivery to cells

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figure 1 from [1] with text explanationWhile many of us are being jabbed in the arm to deliver an agent that stimulates our immune system to recognize the coronavirus SARS-CoV-2 as a threat and destroy it, my research group has been working, in collaboration with colleagues at the European Commission Joint Research Centre, on the dynamics of nanoparticles [1] [see ‘Size matters‘ on October 23rd, 2019] which could be used as carriers for the targeted delivery of therapeutic, diagnostic and imaging agents in the human body [2].  The use of nanoparticles to mechanically stimulate stem cells to activate signalling pathways and modulate their differentiation also has some potential [3]. In studies of the efficacy of nanoparticles in these biomedical applications, the concentration of nanoparticles interacting with the cell is a primary factor influencing both the positive and negative effects.  Such studies often involve exposing a monolayer of cultured cells adhered to the bottom of container to a dose of nanoparticles and monitoring the response over a period of time.  Often, the nominal concentration of the nanoparticles in biological medium supporting the cells is reported and used as the basis for determining the dose-response relationships.  However, we have shown that this approach is inaccurate and leads to misleading results because the nanoparticles in solution are subject to sedimentation due to gravity, Brownian motion [see ‘Slow moving nanoparticles‘ on December 13th, 2017] and inter-particle forces [see ‘ Going against the flow‘ on February 3rd, 2021] which affect their transport within the medium [see graphic] and the resultant concentration adjacent to the monolayer of cells.  Our experimental results using the optical method of caustics [see ‘Holes in fluids‘ on October 22nd, 2014] have shown that nanoparticle size, colloidal stability and solution temperature influence the distribution of nanoparticles in solution.  For particles larger than 60 nm in diameter (about one thousandth of the diameter of a human hair) the nominal dose differs significantly from the dose experienced by the cells.  We have developed and tested a theoretical model that accurately describes the settling dynamics and concentration profile of nanoparticles in solution which can be used to design in vitro experiments and compute dose-response relationships.

References

[1] Giorgi F, Macko P, Curran JM, Whelan M, Worth A & Patterson EA. 2021 Settling dynamics of nanoparticles in simple and biological media. Royal Society Open Science, 8:210068.

[2] Daraee H, Eatemadi A, Abbasi E, Aval SF, Kouhi M, & Akbarzadeh A. 2016 Application of gold nanoparticles in biomedical and drug delivery. Artif. Cells Nanomed. Biotechnol. 44, 410–422. (doi:10.3109/21691401.2014.955107)

[3] Wei M, Li S, & Le W. 2017 Nanomaterials modulate stem cell differentiation: biological
interaction and underlying mechanisms. J. Nanobiotechnol. 15, 75. (doi:10.1186/s12951-
017-0310-5)

Seeing the invisible

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Track of the Brownian motion of a 50 nanometre diameter particle

Track of the Brownian motion of a 50 nanometre diameter particle in a fluid.

Nanoparticles are being used in a myriad of applications including sunscreen creams, sports equipment and even to study the stickiness of snot!  By definition, nanoparticles should have one dimension less than 100 nanometres, which is one thousandth of the thickness of a human hair.  Some nanoparticles are toxic to humans and so scientists are studying the interaction of nanoparticles with human cells.  However, a spherical nanoparticle is smaller than the wavelength length of visible light and so is invisible in a conventional optical microscope used by biologists.  We can view nanoparticles using a scanning electron microscope but the electron beam damages living cells so this is not a good solution.  An alternative is to adjust an optical microscope so that the nanoparticles produce caustics [see post entitled ‘Caustics’ on October 15th, 2014] many times the size of the particle.  These ‘adjustments’ involve closing an aperture to produce a pin-hole source of illumination and introducing a filter that only allows through a narrow band of light wavelengths.  An optical microscope adjusted in this way is called a ‘nanoscope’ and with the addition of a small oscillator on the microscope objective lens can be used to track nanoparticles using the technique described in last week’s post entitled ‘Holes in liquid‘.

The smallest particles that we have managed to observe using this technique were gold particles of diameter 3 nanometres , or about 1o atoms in diameter dispersed in a liquid.

 

Image of 3nm diameter gold particle in a conventional optical microscope (top right), in a nanoscope (bottom right) and composite images in the z-direction of the caustic formed in the nanoscope (left).

Image of 3nm diameter gold particle in a conventional optical microscope (top right), in a nanoscope (bottom right) and composite images in the z-direction of the caustic formed in the nanoscope (left).

Sources:

http://ihcp.jrc.ec.europa.eu/our_activities/nanotechnology/jrc-scientists-develop-a-technique-for-automated-three-dimensional-nanoparticle-tracking-using-a-conventional-microscope

‘Scientists use gold nanoparticles to study the stickiness of snot’ by Rachel Feldman in the Washington Post on October 9th, 2014.

J.-M. Gineste, P. Macko, E.A. Patterson, & M.P. Whelan, Three-dimensional automated nanoparticle tracking using Mie scattering in an optical microscope, Journal of Microscopy, Vol. 243, Pt 2 2011, pp. 172–178

Patterson, E.A., & Whelan, M.P., Optical signatures of small nanoparticles in a conventional microscope, Small, 4(10): 1703-1706, 2008.

Holes in fluids

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Out-of-focus image from optical microscope of 10 micron diameter polystrene spheres in water

Out-of-focus image from optical microscope of 10 micron diameter polystyrene spheres in water

The holes that I wrote about last week and the week before (post entitled ‘Holes‘ on October 8th)were essentially air-filled holes in a solid plate.  When an in-plane load is applied to the plate it deforms and its surface around the hole becomes curved due to the concentration of stress and light passing through the curved surfaces is deviated to form the caustic.  If you didn’t follow that quick recap on last week then you might want flip back to last week’s post before pressing on!

The reverse situation is a solid in a fluid.  It is difficult to induce stress in a fluid so instead we can use a three-dimensional hole, i.e. a sphere, to generate the curve surface for light to pass through and be deviated.  This is quite an easy experiment to do in an optical microscope with some polystyrene spheres floating in distilled water with the microscope slightly out of focus you get bright caustics.  And if you take a series of photographs (the x-y plane) with the microscope objective lens at different heights (z-value) it is possible to reconstruct the three-dimensional shape of the caustic by taking the intensity or greyscale values along the centre line of each image and using them all to create new image of the x-z and, or y-z plane, as shown in the picture.

Well done if you have got this far and are still with me!  I hope you can at least enjoy the pictures.  By the way the particle in the images is about the same diameter as a human hair.

Image in optical microscope of polystrene particle in water (left), series of images at different positions of microscope objective (centre) and artificial image created from greyscale data along centre-lines of image series (right).

Image in optical microscope of polystyrene particle in water (left), series of images at different positions of microscope objective (centre) and artificial image created from greyscale data along centre-lines of image series (right).

Source:

Patterson, E.A., & Whelan, M.P., Tracking nanoparticles in an optical microscope using caustics, Nanotechnology, 19(10): 105502, 2008.