Quantifying nanoscale biochemical heterogeneity in human epithelial cancer cells using combined AFM and PTIR absorption nanoimaging

Kennedy, Eamonn; Al-Majmaie, Rasoul; Al‐Rubeai, Mohamed; Zerulla, Dominic; Rice, James H.

doi:10.1002/jbio.201300138

Cited by 26 publications

(30 citation statements)

References 28 publications

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“…This is consistent both with the PTIR maps reported in ref. 15 on colon adenocarcinoma cells and with the Raman maps taken, in the C-H stretching region where Raman is most sensitive, both on eye lens epithelial cells (LEC) 24 and on HeLa cells. 25 By comparing the AFM maps in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The AFM tip detects the local expansion of the sample when its molecules absorb the radiation to excite their vibrational modes, which occur at frequencies specific to each molecular bond. [7][8][9][10][11] PTIR can thus identify, with a spatial resolution which in biological systems is reported to be on the order of 10 2 nanometers, [12][13][14][15][16] a number of chemical species ranging from nucleic acids to proteins and lipids. The improvement in the sensitivity provided by PTIR with respect to conventional IR microspectroscopy also allows one to study very small quantities of analytes.…”

Section: Introductionmentioning

confidence: 99%

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Protein clustering in chemically stressed HeLa cells studied by infrared nanospectroscopy

et al. 2016

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Photo-Thermal Induced Resonance (PTIR) nanospectroscopy, tuned towards amide-I absorption, was used to study the distribution of proteic material in 34 different HeLa cells, of which 18 were chemically stressed by oxidative stress with NaAsO. The cell nucleus was found to provide a weaker amide-I signal than the surrounding cytoplasm, while the strongest PTIR signal comes from the perinuclear region. AFM topography shows that the cells exposed to oxidative stress undergo a volume reduction with respect to the control cells, through an accumulation of the proteic material around and above the nucleus. This is confirmed by the PTIR maps of the cytoplasm, where the pixels providing a high amide-I signal were identified with a space resolution of ∼300 × 300 nm. By analyzing their distribution with two different statistical procedures we found that the probability to find protein clusters smaller than 0.6 μm in the cytoplasm of stressed HeLa cells is higher by 35% than in the control cells. These results indicate that it is possible to study proteic clustering within single cells by label-free optical nanospectroscopy.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Protein clustering in chemically stressed HeLa cells studied by infrared nanospectroscopy

et al. 2016

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“…Figure shows a series of images of the same position of a bone specimen obtained with different microscopy methods including IR at 1055 cm –1 . A work reported how image analysis applied to a combination of IR absorption nanoimaging and topographic AFM data permitted quantification of chemical complexity at the nanoscale, enabling the analysis of biochemical heterogeneity in mammalian cancer cells on the scale of subcellular features .…”

Section: Infrared Spectroscopymentioning

confidence: 88%

Modern trends in biophotonics for clinical diagnosis and therapy to solve unmet clinical needs

Krafft

2016

Journal of Biophotonics

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This contribution covers recent original research papers in the biophotonics field. The content is organized into main techniques such as multiphoton microscopy, Raman spectroscopy, infrared spectroscopy, optical coherence tomography and photoacoustic tomography, and their applications in the context of fluid, cell, tissue and skin diagnostics. Special attention is paid to vascular and blood flow diagnostics, photothermal and photodynamic therapy, tissue therapy, cell characterization, and biosensors for biomarker detection.

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“…PTIR characterization has been applied to investigate a wide array of samples including bacteria (28,29,(103)(104)(105)(106), cells (107)(108)(109)(110), lipids (111), proteins (112), polymers (30,(113)(114)(115)(116)(117)(118)(119)(120)(121), drugs (122,123), quantum dots (124), plasmonic nanostructures (97,(125)(126)(127), metal-organic frameworks (128), and organo-trihalide perovskites (129). PTIR setups require a pulsed, spectrally narrow, wavelength-tunable laser source; an AFM tip operating in contact mode; and far-field optics to focus light under the tip.…”

Section: Photothermal-induced Resonancementioning

confidence: 99%

Infrared Imaging and Spectroscopy Beyond the Diffraction Limit

Centrone

2015

Annual Rev. Anal. Chem.

243

275

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Progress in nanotechnology is enabled by and dependent on the availability of measurement methods with spatial resolution commensurate with nanomaterials' length scales. Chemical imaging techniques, such as scattering scanning near-field optical microscopy (s-SNOM) and photothermal-induced resonance (PTIR), have provided scientists with means of extracting rich chemical and structural information with nanoscale resolution. This review presents some basics of infrared spectroscopy and microscopy, followed by detailed descriptions of s-SNOM and PTIR working principles. Nanoscale spectra are compared with far-field macroscale spectra, which are widely used for chemical identification. Selected examples illustrate either technical aspects of the measurements or applications in materials science. Central to this review is the ability to record nanoscale infrared spectra because, although chemical maps enable immediate visualization, the spectra provide information to interpret the images and characterize the sample. The growing breadth of nanomaterials and biological applications suggest rapid growth for this field.

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Quantifying nanoscale biochemical heterogeneity in human epithelial cancer cells using combined AFM and PTIR absorption nanoimaging

Abstract: Rice, J.H. (2015) "Quantifying nanoscale biochemical heterogeneity in human epithelial cancer cells using combined AFM and PTIR absorption nanoimaging" Journal of Biophotonics, 8(1-2) : 133-141 which has been published in final form at http://dx

Cited by 26 publications

References 28 publications

Protein clustering in chemically stressed HeLa cells studied by infrared nanospectroscopy

Protein clustering in chemically stressed HeLa cells studied by infrared nanospectroscopy

Modern trends in biophotonics for clinical diagnosis and therapy to solve unmet clinical needs

Infrared Imaging and Spectroscopy Beyond the Diffraction Limit

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