A Novel, All-Optical Tool for Controllable and Non-Destructive Poration of Cells with Single-Micron Resolution

Casey, Duncan; Wylie, Douglas; Gallo, Juan; Dent, Michael R.; Salehi-Reyhani, Ali; Wilson, Rab; Brooks, Nicholas J.; Long, N.J.; Willison, Keith R.; Klug, David R.; Neil, Mark A. A.; Neale, Steven L.; Cooper, Jonathan M.; Ces, Oscar

doi:10.1364/boda.2015.bw1a.5

Cited by 6 publications

(4 citation statements)

References 13 publications

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“…Such techniques are well-covered elsewhere in this book and will not be covered further here, although research by the groups of Jon Cooper and Steven Neale in Glasgow have recently demonstrated that such a platform can be utilised to electroporate cells selecting both spatially and for cell morphology, based upon the "electrochemical shadow" that the cells cast upon the amorphous silicon surface [3,70]. Amorphous silicon layers can be used to directly facilitate optoporation, however: the microsecond-pulse experiments of Fan et al [66,71] and continuous-wave experiments in our own laboratories [72] have demonstrated that hot-spot generation upon infrared irradiation of thin-layer silicon is sufficient to cause singleor even sub-cellular poration (see Figure 9, below).…”

Section: 32mentioning

confidence: 99%

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Optical Tools for Single-Cell Manipulation and Analysis

Casey

Dooley

2016

Series in BioEngineering

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Experiments on individual cells require a range of extremely precise tools to permit their selection, manipulation, stimulation and analysis. This is further complicated by the cells' sensitivity to their environment, meaning that such tools must also be very gentle (or at least very localised) to minimise the generation of artefacts. Optical tools provide ideal performance in a number of such roles, exhibiting high spatial and temporal selectivity while causing minimal non-specific effects. This review will focus upon the optical tools that have been developed for these purposes, ranging from optical trapping systems which provide a contact-free technique for the manipulation of micron-scale objects, through to a selection of the different optically-mediated cell membrane disruption methods available for lysis and/or delivery of material.Optical trapping techniques provide the means to manipulate matter at approximately a 1-100 µm scale, without requiring direct contact with the cell [1]. The use of infrared wavelengths minimises the amount of light absorbed by biological targets, while a range of light-sculpting approaches are available to generate a wide array of complex beams which can be dynamically modified. This means that cells (or micro-structured probes) can be directly manipulated, either to build arrays or to perform mechanical measurements of the properties of the cell membrane.However, to modulate or measure processes occurring beyond the cell membrane, a mechanism of controlled membrane rupture must be utilised. These can either be destructive, for selective lysis experiments, or reversible, to allow the introduction of material to stimulate responses with minimal disruption to the cell. Both approaches feature a range of modalities: for example, lysis can be induced by direct plasma formation using high-energy pulsed lasers to induce catastrophic damage to anything within a defined radius [2], or can be combined with electrical fields to provide lysis with single-cell resolution [3]. Similarly, photoporation can be accomplished directly with very high precision (although conditions must be finely tuned to minimise cell disruption), or in combination with materials with specific absorption characteristics to deliver similar effects using much longer wavelengths and commensurately lower cell damage [4].The theories underpinning these techniques will be discussed, and illuminated using examples of recent research to provide first-hand examples of their successful application. The advantages and disadvantages of each approach will be comprehensively debated, and directions of promising research will be presented to give insight into the tools and techniques likely to be available in the future.

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Section: 32mentioning

confidence: 99%

“…HCT116 cells [73] were cultured directly onto thin-layer amorphous silicon and assembled into a closed chip, then inverted so as to hang pendant. (Below) Bright-field and fluorescence images showing selective poration of HCT cells, illustrated using propidium iodide [72].…”

Section: 32mentioning

confidence: 99%

Optical Tools for Single-Cell Manipulation and Analysis

Casey

Dooley

2016

Series in BioEngineering

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“…We found that cells grown on amorphous silicon (as used in solar panels) selectively and reversibly developed membrane nanopores under illumination from IR lasers, to which they would typically be transparent. 11 We can therefore deliver materials (such as dyes, nucleic acid, and proteins) to specific cells and even to specific regions of a single cell, in a way that does not obviously damage the cell's ability to function (see Figure 2). Observations like these have previously been made with different substrates, 12 but we were able to measure the pores' diameters using quantum dots as sizing tools, and thus provide wholly new insights into the pores' energetics and their effects on the target cell.…”

Section: 1117/21201605006474 Page 2/3mentioning

confidence: 99%

Laser-based techniques for non-destructive surgery on individual cells

Casey¹,

Wylie²

2016

SPIE Newsroom

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“…In addition, success rates were low. More recently, our own work has identified optothermally-produced pores with diameters of ≤10 nm measured using quantum dots, but the mechanism behind their formation remains open to discussion (Casey et al 2015).…”

Section: Mechanism(s) Of Porationmentioning

confidence: 99%

Optically-controlled platforms for transfection and single- and sub-cellular surgery

2015

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Improving the resolution of biological research to the single-cell or sub-cellular level is of critical importance in a wide variety of processes and disease conditions. Most obvious are those linked to aging and cancer, many of which are dependent upon stochastic processes where individual, unpredictable failures or mutations in individual cells can lead to serious downstream conditions across the whole organism. The traditional tools of biochemistry struggle to observe such processes: the vast majority are based upon ensemble approaches analysing the properties of bulk populations, which means that details of individual constituents is lost. What are required, then, are tools with the precision and resolution to probe and dissect cells at the single-micron scale: the scale of the individual organelles and structures that control their function. In this review, we highlight the use of highly-focused laser beams to create systems which provide precise control and specificity at the single-cell or even single-micron level. The intense focal points generated can directly interact with cells and cell membranes, which in conjunction with related modalities such as optical trapping provide a broad platform for the development of single-cell and sub-cellular surgery approaches. These highly tuneable tools have been demonstrated to deliver or remove material from cells of interest, and they can simultaneously excite fluorescent probes for imaging purposes or plasmonic structures for very local heating. We discuss both the history and recent applications of the field, highlighting the key findings and developments over the last 40 years of biophotonics research.

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A Novel, All-Optical Tool for Controllable and Non-Destructive Poration of Cells with Single-Micron Resolution

Cited by 6 publications

References 13 publications

Optical Tools for Single-Cell Manipulation and Analysis

Optical Tools for Single-Cell Manipulation and Analysis

Laser-based techniques for non-destructive surgery on individual cells

Optically-controlled platforms for transfection and single- and sub-cellular surgery

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