Non-invasive label-free monitoring the cardiac differentiation of human embryonic stem cells in-vitro by Raman spectroscopy

Pascut, Flavius C.; Kalra, Spandan; George, Vinoj; Welch, Nathan; Denning, Chris; Notingher, Ioan

doi:10.1016/j.bbagen.2013.01.030

Cited by 68 publications

(70 citation statements)

References 30 publications

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“…[9][10][11][12][13] The integration of environmental chambers with inverted Raman microscopes can allow time-and spatially-resolved molecular studies of cellular processes spanning days and weeks, such as stem cell differentiation. 14,15 Nevertheless, RMS has been used only in few studies to characterize molecular details associated with host-parasite interaction. For example, RMS has been utilized to study the protozoan pathogen Plasmodium falciparum, the agent of human malaria.…”

Section: Introductionmentioning

confidence: 99%

Analysis of interaction between the apicomplexan protozoan Toxoplasma gondii and host cells using label-free Raman spectroscopy

Naemat

Elsheikha

Al-sandaqchi

et al. 2015

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Label-free imaging using Raman micro-spectroscopy (RMS) was used to characterize the spatio-temporal molecular changes of T. gondii tachyzoites and their host cell microenvironment. Raman spectral maps were recorded from isolated T. gondii tachyzoites and T. gondii-infected human retinal cells at 6 hr, 24 hr and 48 hr postinfection. Principal component analysis (PCA) of the Raman spectra of paraformaldehyde-fixed infected cells indicated a significant increase in the amount of lipids and proteins in the T. gondii tachyzoites as the infection progresses within host cells. These results were confirmed by experiments carried out on live T. gondii-infected cells and were correlated with an increase in the concentration of proteins and lipids required for the replication of this intracellular pathogen. These findings demonstrate the potential of RMS to characterize time-and spatially-dependent molecular interactions between intracellular pathogens and the host cells. Such information may be useful for discovery of pharmacological targets or screening compounds with potential neuroprotective activity for eminent effects of changes in brain infection control practices. 2

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Section: Introductionmentioning

confidence: 99%

Analysis of interaction between the apicomplexan protozoan Toxoplasma gondii and host cells using label-free Raman spectroscopy

Naemat

Elsheikha

Al-sandaqchi

et al. 2015

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“…Because Raman spectroscopy is non-labeling and non-destructive, repeated measurements can be made of the same sample. This allows time-lapsed traces of biological proceses [136,145].…”

Section: Applicationsmentioning

confidence: 99%

“…This microscope is equipped with an environmental chamber allowing live cells to be studied over several days. Raman spectroscopy is well-suited to study biological processes occuring on the order of several minutes to a few days including cell-cell interactions [133], apoptosis [134], mitosis [135], and stem cell differentiation [136].…”

Section: Typical Systemmentioning

confidence: 99%

Raman spectroscopy: techniques and applications in the life sciences

Shipp¹,

Sinjab²,

Notingher³

2017

Adv. Opt. Photon.

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189

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Raman spectroscopy is an increasingly popular technique in many areas including biology and medicine.It is based on Raman scattering, a phenomenon in which incident photons lose or gain energy via interactions with vibrating molecules in a sample. These energy shifts can be used to obtain information regarding molecular composition of the sample with very high accuracy. Applications of Raman spectroscopy in the life sciences have included quantification of biomolecules, hyperspectral molecular imaging of cells and tissue, medical diagnosis, and others. This review briefly presents the physical origin of Raman scattering explaining the key classical and quantum mechanical concepts. Variations of the Raman effect will also be considered, including resonance, coherent, and enhanced Raman scattering. We discuss the molecular origins of prominent bands often found in the Raman spectra of biological samples. Finally, we examine several variations of Raman spectroscopy techniques in practice, looking at their applications, strengths, and challenges. This review is intended to be a starting resource for scientists new to Raman spectroscopy, providing theoretical background and practical examples as the foundation for further study and exploration.

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“…Raman spectroscopy (RS) is a well-established analytical tool for measuring non-invasively live cells [3][4][5][6] and bone tissue [7][8][9] . RS has been used to measure the deposition of hydroxyapatite (HA) (the mineral phase of bone) on various types of bone tissue engineering scaffolds 10,11 and characterize the mineralization of osteoblast cultures in-vitro [12][13][14][15] .…”

Section: Introductionmentioning

confidence: 99%

Feasibility of Spatially Offset Raman Spectroscopy for in Vitro and in Vivo Monitoring Mineralization of Bone Tissue Engineering Scaffolds

et al. 2016

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We investigated the feasibility of using spatially offset Raman spectroscopy (SORS) for nondestructive characterization of bone tissue engineering scaffolds. The deep regions of these scaffolds, or scaffolds implanted subcutaneously in live animals, are typically difficult to measure by confocal Raman spectroscopy techniques because of the limited depth penetration of light caused by the high level of light scattering. Layered samples consisting of bioactive glass foams (IEIC16), three-dimensional (3D)-printed biodegradable poly(lactic-co-glycolic acid) scaffolds (PLGA), and hydroxyapatite powder (HA) were used to mimic nondestructive detection of biomineralization for intact real-size 3D tissue engineering constructs. SORS spectra were measured with a new SORS instrument using a digital micromirror device (DMD) to allow software selection of the spatial offsets. The results show that HA can be reliably detected at depths of 0–2.3 mm, which corresponds to the maximum accessible spatial offset of the current instrument. The intensity ratio of Raman bands associated with the scaffolds and HA with the spatial offset depended on the depth at which HA was located. Furthermore, we show the feasibility for in vivo monitoring mineralization of scaffold implanted subcutaneously by demonstrating the ability to measure transcutaneously Raman signals of the scaffolds and HA (fresh chicken skin used as a top layer). The ability to measure spectral depth profiles at high speed (5 s acquisition time) and the ease of implementation make SORS a promising approach for noninvasive characterization of cell/tissue development in vitro, and for long-term in vivo monitoring the mineralization in 3D scaffolds subcutaneously implanted in small animals.

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Non-invasive label-free monitoring the cardiac differentiation of human embryonic stem cells in-vitro by Raman spectroscopy

Cited by 68 publications

References 30 publications

Analysis of interaction between the apicomplexan protozoan Toxoplasma gondii and host cells using label-free Raman spectroscopy

Analysis of interaction between the apicomplexan protozoan Toxoplasma gondii and host cells using label-free Raman spectroscopy

Raman spectroscopy: techniques and applications in the life sciences

Feasibility of Spatially Offset Raman Spectroscopy for in Vitro and in Vivo Monitoring Mineralization of Bone Tissue Engineering Scaffolds

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