Evidence of Pressure Waves in Confined Laser Ablation

Domke, Matthias; Felsl, Dominik; Rapp, Stephan; Sotrop, Jürgen; Huber, Heinz P.; Schmidt, Michael; Ressel, Josef; Erlangen-Nürnberg, Friedrich-Alexander-Universität

doi:10.2961/jlmn.2015.02.0002

Cited by 10 publications

(6 citation statements)

References 1 publication

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“…As for the physical mechanism of ejection, it depends on a few parameters, such as laser pulse energy, laser pulse duration, and thickness of the film. In previous reports, it was found that the shock wave produced the force to push the cells when the laser pulse duration was at the level of picosecond or femtosecond ( 31 , 32 ). When the pulse duration was at the level of nanosecond or longer, the gas pressure was dominated ( 25 – 30 ).…”

Section: Discussionmentioning

confidence: 99%

“…The ejection mechanism of LIFT is dependent on thickness of the coating layer, material, and laser power and duration, but the basic setup is similar ( 24 ). When a laser pulse shines on the surface of the coating layer, it will absorb the laser energy and vaporize, and the material on it will be pushed away under the gas pressure ( 25 – 30 ) or shock wave ( 31 , 32 ) induced by heating.…”

Section: Introductionmentioning

confidence: 99%

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Isolation and Culture of Single Microbial Cells by Laser Ejection Sorting Technology

Peng

Liu

Wang³

et al. 2022

Appl Environ Microbiol

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Single cell isolation and cultivation play an important role in studying physiology, gene expression and functions of microorganisms. A series of single-cell isolation technologies have been developed, among which single-cell ejection technology is one of the most promising. Single cell ejection technology has applied Laser Induced Forward Transfer Technique (LIFT) to isolate bacteria but the viability (or recovery rate) of cells after sorting has not been clarified in the current research progress. In this work, to keep the cells alive as much as possible, we propose a three-layer LIFT system (top layer: 25-nm aluminum film; second layer: 3 μm agar media; third layer: liquid containing bacterial) for the isolation and cultivation of single Gram-negative ( E. coli ), Gram-positive ( Lactobacillus rhamnosus GG, LGG), and eukaryotic microorganisms ( Saccharomyces cerevisiae ). The experiment results showed that the average survival rates for ejected pure single cells were 63% for Saccharomyces cerevisiae , 22% for E. coli DH5α, and 74% for LGG. In addition, we successfully isolated and cultured the GFP expressing E. coli JM109 from the mixture containing complex communities of soil bacteria by fluorescence signal. The average survival rate of E. coli JM109 was demonstrated to be 25.3%. In this study, the isolated and cultured single colonies were further confirmed by colony PCR and sequencing. Such precise sorting and cultivation technique of live single microbial cells could be coupled with other microscopic approaches to isolate single microorganisms with specific functions, revealing their roles in the natural community. Importance We developed a laser induced forward transfer (LIFT) technology to accurately isolate single live microbial cells. The cultivation recovery rates of the ejected single cells were 63% for Saccharomyces cerevisiae , 22% for E. coli DH5α, and 74% for Lactobacillus rhamnosus GG (LGG). Coupled LIFT with fluorescent microscope, we demonstrated that single cells of GFP expressing E. coli JM109 were sorted according to fluorescence signal from a complex community of soil bacteria, and subsequently cultured with 25% cultivation recovery rate. This single cell live sorting technology could isolate single microbes with specific functions, revealing their roles in the natural community.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Isolation and Culture of Single Microbial Cells by Laser Ejection Sorting Technology

Peng

Liu

Wang³

et al. 2022

Appl Environ Microbiol

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“…An energy absorption layer is coated on a transparent substrate which we called donor, and then the material to be printed is spread onto the donor. When a laser pulse propagates through the transparent substrate and focuses onto the surface of the energy absorption layer, the energy absorption layer will be heated to vaporize, and the material on it will be pushed away under the gas pressure (22)(23)(24)(25)(26)(27) or the shock wave (28,29) induced by heating.…”

Section: Introductionmentioning

confidence: 99%

Laser induced isolation and cultivation of single microbial cells

Peng

Wang

Wang³

et al. 2020

Preprint

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Single cell isolation and cultivation play an important role in studying physiology, gene expression and functions of microorganisms. Laser Induced Forward Transfer Technique (LIFT) has been applied to isolate single cells but the cell viability after sorting is unclear. We demonstrate that a three-layer LIFT system could be applied to isolate single cells of Gram-negative (E. coli), Gram-positive (Lactobacillus rhamnosus GG, LGG), and eukaryotic microorganisms (Saccharomyces cerevisiae) and the sorted single cells were able to be cultured. The experiment results showed that the average cultivation recovery rate of the ejected single cells were 58% for Saccharomyces cerevisiae, 22% for E. coli, and 74% for Lactobacillus rhamnosus GG (LGG). The identities of the cultured cells from single cell sorting were confirmed by using colony PCR with 16S-rRNA for bacteria and large unit rRNA for yeast and subsequent sequencing. This precise sorting and cultivation technique of live single microbial cells can be coupled with other microscopic approaches (e.g. fluorescent and Raman microscopy) to culture single microorganisms with specific functions, revealing their roles in the natural community.ImportanceSingle cell isolation and cultivation are crucial to recover microorganisms for the study of physiology, gene expression and functions. We developed a laser induced cell sorting technology to precisely isolate single microbial cells from a microscopic slide. More importantly, the isolated single microbial cells are still viable for cultivation. We demonstrate to apply the live sorting method to isolate and cultivate single cells of Gram-negative (E. coli), Gram-positive (Lactobacillus rhamnosus GG, LGG), and eukaryotic microorganisms (Saccharomyces cerevisiae). This precise sorting and cultivation technique can be coupled with other microscopic approaches (e.g. fluorescent and Raman microscopy) to culture specifically targeted single microorganisms from microbial community.Abstract Graphic

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“…During experiments, we observed bright spots remaining on the coverslip after ejection ( Figure 2C ). The spectra of these bright spots ( Figure S4C ) were distinguishable from those of PS or PMMA ( Figure S4A ), indicating that these bright spots might have been caused by the deformations in the TiO 2 coating (45, 46).…”

Section: Resultsmentioning

confidence: 99%

High-throughput single-cell sorting by stimulated Raman-activated cell ejection

Zhang,

Lin,

et al. 2023

Preprint

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Single-cell sorting is essential to explore cellular heterogeneity in biology and medicine. Recently developed Raman-activated cell sorting (RACS) circumvents the limitations of fluorescence-activated cell sorting, such as the cytotoxicity of labels. However, the sorting throughputs of all forms of RACS are limited by the intrinsically small cross-section of spontaneous Raman scattering. Here, we report a stimulated Raman-activated cell ejection (S-RACE) platform that enables high-throughput single-cell sorting based on high-resolution multi-channel stimulated Raman chemical imaging, in situ image decomposition, and laser-induced cell ejection. The performance of this platform was illustrated by sorting a mixture of 1 μm polymer beads, where 95% yield, 98% purity, and 14 events per second throughput were achieved. Notably, our platform allows live cell ejection, allowing for the growth of single colonies of bacteria and fungi after sorting. To further illustrate the chemical selectivity, lipid-rich Rhodotorula glutinis cells were successfully sorted from a mixture with Saccharomyces cerevisiae, confirmed by downstream quantitative PCR. Furthermore, by integrating a closed-loop feedback control circuit into the system, we realized real-time single-cell imaging and sorting, and applied this method to precisely eject regions of interest from a rat brain tissue section. The reported S-RACE platform opens exciting opportunities for a wide range of single-cell applications in biology and medicine.

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Evidence of Pressure Waves in Confined Laser Ablation

Cited by 10 publications

References 1 publication

Isolation and Culture of Single Microbial Cells by Laser Ejection Sorting Technology

Isolation and Culture of Single Microbial Cells by Laser Ejection Sorting Technology

Laser induced isolation and cultivation of single microbial cells

High-throughput single-cell sorting by stimulated Raman-activated cell ejection

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