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Campagnola, Paul J.; Millard, Andrew C.; Terasaki, Mark; Hoppe, Pamela E.; Malone, Christian J.; Mohler, William A.

doi:10.1016/j.bpj.2012.07.025

Cited by 29 publications

(31 citation statements)

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“…Collagen, due to its helical nature, is a noncentrosymmetric structure. It exhibits inherently strong second‐harmonic generation (SHG) . The propensity of a material to generate SHG can be described by its second‐order nonlinear susceptibility tensor χ (2) .…”

Section: Introductionmentioning

confidence: 99%

Collagen chirality and three‐dimensional orientation studied with polarimetric second‐harmonic generation microscopy

Golaraei

Mirsanaye

et al. 2018

Journal of Biophotonics

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Polarization‐dependent second‐harmonic generation (P‐SHG) microscopy is used to characterize molecular nonlinear optical properties of collagen and determine a three‐dimensional (3D) orientation map of collagen fibers within a pig tendon. C6 symmetry is used to determine the nonlinear susceptibility tensor components ratios in the molecular frame of reference χzzz2/χzxx2 and χxyz2/χzxx2, where the latter is a newly extracted parameter from the P‐SHG images and is related to the chiral structure of collagen. The χxyz2/χzxx2 is observed for collagen fibers tilted out of the image plane, and can have positive or negative values, revealing the relative polarity of collagen fibers within the tissue. The P‐SHG imaging was performed using a linear polarization‐in polarization‐out (PIPO) method on thin sections of pig tendon cut at different angles. The nonlinear chiral properties of collagen can be used to construct the 3D organization of collagen in the tissue and determine the orientation‐independent molecular susceptibility ratios of collagen fibers in the molecular frame of reference.

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Section: Introductionmentioning

confidence: 99%

Collagen chirality and three‐dimensional orientation studied with polarimetric second‐harmonic generation microscopy

Golaraei

Mirsanaye

et al. 2018

Journal of Biophotonics

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“…In the last few years there have been a number of applications of nonlinear microscopy to cartilage. Second harmonic (SHG) imaging of collagen is well established [5,6]. Its application to cartilage is complicated by the fact that the individual fibres of type II collagen are finer than those of type I and below the limit of resolution, but valuable information on its organisation in cartilage has been derived by image processing [7,8] and polarization senstive measurements [9].…”

Section: Introductionmentioning

confidence: 99%

Chemically specific imaging and in-situ chemical analysis of articular cartilage with stimulated Raman scattering

et al. 2013

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Stimulated Raman Scattering has been applied to unstained samples of articular cartilage enabling the investigation of living cells within fresh tissue. Hyperspectral SRS measurements over the CH vibrational region showed variations in protein and lipid content within the cells, pericellular matrix and interteritorial matrix. Changes in the cells and pericellular matrix were investigated as a function of depth into the cartilage. Lipid was detected in the pericellular matrix of superficial zone chondrocytes. The spectral profile of lipid droplets within the chondrocytes indicated that they contained predominantly unsaturated lipids. The mineral content has been imaged by using the PO4 3-vibration at 959cm -1 and the CO3 2-vibration at 1070cm -1 . Both changes in cells and mineralization are known to be important factors in the progression of osteoarthritis. SRS enables these to be visualised in fresh unstained tissue and consequently should benefit osteoarthiritis research

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“…A more powerful imaging modality is represented by two-photon excited fluorescence (TPF) microscopy [20], which is a high-resolution laser scanning imaging technique enabling deep optical imaging of tissues, as demonstrated by studies performed on ex vivo tissue samples [21][22][23][24][25], fresh biopsies [26][27][28][29][30][31], and also in vivo on both animals [32] and humans [33][34][35][36][37][38]. Additional morphological information can be provided by second-harmonic generation (SHG) microscopy [39][40][41][42][43][44][45][46][47][48][49], which can be combined with TPF microscopy using the same laser source. Combined TPF-SHG microscopy represents a powerful tool for imaging skin dermis, since the main dermal components, collagen and elastin, can be imaged by SHG and TPF microscopy, respectively [22].…”

Section: Introductionmentioning

confidence: 99%

In vivo non‐invasive monitoring of collagen remodelling by two‐photon microscopy after micro‐ablative fractional laser resurfacing

Cicchi

Kapsokalyvas²,

Troiano

et al. 2013

Journal of Biophotonics

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Journal of BIOPHOTONICSNon-linear optical microscopy is becoming popular as a non-invasive in vivo imaging modality in dermatology. In this study, combined TPF and SHG microscopy were used to monitor collagen remodelling in vivo after microablative fractional laser resurfacing. Papillary dermis of living subjects, covering a wide age range, was imaged immediately before and forty days after treatment. A qualitative visual examination of acquired images demonstrated an age-dependent remodelling effect on collagen. Additional quantitative analysis of new collagen production was performed by means of two image analysis methods. A higher increase in SHG to TPF ratio, corresponding to a stronger treatment effectiveness, was found in older subjects, whereas the effect was found to be negligible in young, and minimal in middle age subjects. Analysis of collagen images also showed a dependence of the treatment effectiveness with age but with controversial results. While the diagnostic potential of in vivo multiphoton microscopy has already been demonstrated for skin cancer and other skin diseases, here we first successfully explore its potential use for a non-invasive follow-up of a laser-based treatment.Three-dimensional projection of a stack of 20 SHG images acquired within dermis of a healthy subject. Volume shown: 400 Â 400 Â 100 mm 3 .

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Cited by 29 publications

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Collagen chirality and three‐dimensional orientation studied with polarimetric second‐harmonic generation microscopy

Collagen chirality and three‐dimensional orientation studied with polarimetric second‐harmonic generation microscopy

Chemically specific imaging and in-situ chemical analysis of articular cartilage with stimulated Raman scattering

In vivo non‐invasive monitoring of collagen remodelling by two‐photon microscopy after micro‐ablative fractional laser resurfacing

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