Broadband Fourier-transform coherent Raman spectroscopy with an ytterbium fiber laser

Coluccelli, Nicola; Vicentini, Edoardo; Alessio, Giovanni; Howle, Christopher R.; McEwan, Ken; Laporta, P.; Galzerano, G.

doi:10.1364/oe.26.018855

Cited by 12 publications

(12 citation statements)

References 17 publications

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“…The system was based on a single high-power Ybfiber laser source coupled to a FT interferometer with pixel dwell time of 160 μs (~675 spectral data points). 134 Thus, such studies focused on development of rapid CRS imaging techniques demonstrate the promise of coupling high-content spectral imaging with highthroughput single-cell analysis.…”

Section: High-speed Crs Imaging-microfluidic Devices Have Enabled Resmentioning

confidence: 99%

Quantitative imaging of lipid droplets in single cells

Gupta

Dorlhiac

Streets

2019

Analyst

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Section: High-speed Crs Imaging-microfluidic Devices Have Enabled Resmentioning

confidence: 99%

Quantitative imaging of lipid droplets in single cells

Gupta

Dorlhiac

Streets

2019

Analyst

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“…Therefore, a fiber laser-based FT-CARS implementation would lower the cost and size of the apparatus and may be the best direction for future implementations. [66] In closing, we wish to highlight the need for a uniform metric in reporting sorting performance in microfluidic-based RACS machines. Particularly, the relationship between purity and yield means that reporting only purity or only yield for a single experiment is an insufficient metric for judging performance.…”

Section: Discussionmentioning

confidence: 99%

High‐Throughput Raman‐Activated Cell Sorting in the Fingerprint Region

Lindley

Pablo

Peterson

et al. 2022

Adv Materials Technologies

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Cell sorting is the workhorse of biological research and medicine. [1] Cell sorters are commonly used to sort heterogeneous populations of cells based on their intrinsic features and have numerous applications in microbiology, immunology, virology, stem cell biology, and metabolic engineering. [2][3][4][5][6] While several schemes have been developed for sorting cells based on features such as size, [7] morphology, [8][9][10] viability, [11,12] and surface antigens, [13] those able to sort at high throughput for intracellular chemical content are limited to fluorescence-activated and Raman-activated cell sorting (FACS and RACS, respectively). [1,14] In both methods, sorting is accomplished by a stepwise automatic process: 1) measurement of chemical content with single-cell resolution, 2) analysis of the measurement data to identify cells of interest, and 3) mechanical separation of target cells from the original sample. [1] FACS machines commonly identify molecular targets by signals from attached exogenous fluorophores or coexpressed transgenic fluorescent proteins. This indirect detection scheme is often referred to as labeled detection. [1] Although fluorescent labeling is essential for microbiological research, the use of labels has inherrent drawbacks. These include reduced cellular vitality due to the cytotoxicity of labels themselves, nonspecific binding of labels, a lack of labels or transfection methods for many molecular targets and species, incompatibility with human-targeted cell therapies, and increased experimental complexity imposed by labeling protocols. [14] On the other hand, RACS machines directly identify target molecules in a label-free manner via Raman spectroscopy, which measures the inelastic scattering of incident photons by characteristic molecular vibrations. [14] The chemical specificity of this optical measurement contrasts with non-chemically-specific label-free methods such as impedance-based sorting for cell viability and image-based sorting for morphology or deformability. [10][11][12] To date, FACS technology is well developed, commercialized, and broadly adopted, while RACS is nascent and comprised of a small number of lab-based demonstrations. [14][15][16][17][18][19][20][21][22][23][24][25][26] A major hurdle in the broad deployment of RACS for biomedical applications is the weak signal produced by the Raman scattering process. This necessitates comparatively Cell sorting is the workhorse of biological research and medicine. Cell sorters are commonly used to sort heterogeneous cell populations based on their intrinsic features. Raman-activated cell sorting (RACS) has recently received considerable interest by virtue of its ability to discriminate cells by their intracellular chemical content, in a label-free manner. However, the broad deployment of RACS beyond lab-based demonstrations is hindered by a fundamental trade-off between throughput and measurement bandwidth (i.e., cellular information content). Here this trade-off is overcome and broadband RACS in the fingerprint reg...

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“…The copyright holder for this preprint (which this version posted May 17, 2021. ; https://doi.org/10.1101/2021.05.16.444384 doi: bioRxiv preprint a challenge to widespread adoption. Therefore, a fiber laser-based FT-CARS implementation 55 would lower the cost and size of the apparatus.…”

Section: Discussionmentioning

confidence: 99%

“…Third, the Ti:Sapphire pulsed laser and optics used in this system constitute a considerable expense that presents a challenge to widespread adoption. Therefore, a fiber laser-based FT-CARS implementation 55 would lower the cost and size of the apparatus.…”

Section: Discussionmentioning

confidence: 99%

High-throughput Raman-activated cell sorting in the fingerprint region

Lindley

et al. 2021

Preprint

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Cell sorting is the workhorse of biological research and medicine. Cell sorters are commonly used to sort heterogeneous cell populations based on their intrinsic features. Raman-activated cell sorting (RACS) has recently received considerable interest by virtue of its ability to discriminate cells by their intracellular chemical content, in a label-free manner. However, broad deployment of RACS beyond lab-based demonstrations is hindered by a fundamental trade-off between throughput and measurement bandwidth (i.e., cellular information content). Here we overcome this trade-off and demonstrate broadband RACS in the fingerprint region (300 - 1,600 cm-1) with a record high throughput of ~50 cells per second. This represents a 100x throughput increase compared to previous demonstrations of broadband fingerprint-region RACS. To show the utility of our RACS, we demonstrate real-time label-free sorting of microalgal cells based on their accumulation of carotenoids and polysaccharide granules. These results hold promise for medical, biofuel, and bioplastic applications.

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Broadband Fourier-transform coherent Raman spectroscopy with an ytterbium fiber laser

Cited by 12 publications

References 17 publications

Quantitative imaging of lipid droplets in single cells

Quantitative imaging of lipid droplets in single cells

High‐Throughput Raman‐Activated Cell Sorting in the Fingerprint Region

High-throughput Raman-activated cell sorting in the fingerprint region

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