Investigation into the structure of crystalline maltodextrin particles by second harmonic generation microscopy

Bennett, Elisha; Harvey, MacAulay; Cisek, Richard; Tokarz, Danielle

doi:10.1364/boe.481689

Cited by 5 publications

(2 citation statements)

References 30 publications

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“…PIPO-SHG imaging was performed using a custom-built laser scanning microscope, previously described in [ 21 , 22 ] and is schematically shown in Fig. 1(a) .…”

Section: Methodsmentioning

confidence: 99%

Effect of out of plane orientation on polarization second harmonic generation of single collagen fibrils

Harvey,

Cisek,

Tokarz

et al. 2023

Biomed. Opt. Express

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Second harmonic generation (SHG) microscopy has emerged as a powerful technique for visualizing collagen organization within tissues. Amongst the many advantages of SHG is its sensitivity to collagen nanoscale organization, and its presumed sensitivity to the relative out of plane polarity of fibrils. Recent results have shown that circular dichroism SHG (CD-SHG), a technique that has been commonly assumed to reveal the relative out of plane polarity of collagen fibrils, is actually insensitive to changes in fibril polarity. However, results from another research group seem to contradict this conclusion. Both previous results have been based on SHG imaging of collagen fibrils within tissues, therefore, to gain a definitive understanding of the sensitivity of SHG to relative out of plane polarity, the results from individual fibrils are desirable. Here we present polarization resolved SHG microscopy (PSHG) data from individual collagen fibrils oriented out of the image plane by buckling on an elastic substrate. We show through correlation with atomic force microscopy measurements that SHG intensity can be used to estimate the out of plane angle of individual fibrils. We then compare the sensitivity of two PSHG techniques, CD-SHG and polarization-in, polarization-out SHG (PIPO-SHG), to the relative out of plane polarity of individual fibrils. We find that for single fibrils CD-SHG is insensitive to relative out of polarity and we also demonstrate the first direct experimental confirmation that PIPO-SHG reveals the relative out of plane polarity of individual collagen fibrils.

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“…PIPO-SHG imaging was performed using a custom-built laser scanning microscope, previously described in [ 21 , 22 ] and is schematically shown in Fig. 1(a) .…”

Section: Methodsmentioning

confidence: 99%

Effect of out of plane orientation on polarization second harmonic generation of single collagen fibrils

Harvey,

Cisek,

Tokarz

et al. 2023

Biomed. Opt. Express

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“…PSHG imaging is performed using a custom-built laser scanning microscope previously described in [15,16] (schematically shown in Fig. 1a) coupled to an ultrafast laser (FemtoLux 3, EKSPLA) with a center wavelength of 1030 nm, pulse duration of ~290 fs, and repetition rate of 5 MHz.…”

Section: Microscope Setupmentioning

confidence: 99%

Determining the helical pitch and relative polarity of collagen using polarization-resolved SHG

Harvey,

Cisek,

Kreplak

et al. 2024

Multiphoton Microscopy in the Biomedical Sciences XXIV

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Collagen fibrils are nanoscale biological ropes which are the primary load bearing structures within the human body. Due to the non-centrosymmetric structure and high nonlinearity of the triple helical collagen molecule, collagen fibrils are able to produce significant second harmonic generation (SHG) when placed at the focus of an ultrafast laser. There have been many publications which have utilized polarization resolved SHG (PSHG) for investigating collagen organization in tissues, mainly for applications in disease diagnosis. However so far there has been little work on investigating individual isolated collagen fibrils using PSHG. This presents an exciting opportunity for research since PSHG imaging of individual fibrils can help us to gain a better understanding of PSHG images of collections of fibrils in tissues, and therefore improve the utility of PSHG as a potential diagnostic tool. Additionally, the high sensitivity of PSHG to changes in molecular structure can provide us with a better understanding of the nanoscale origins of injuries to tissues such as skin, tendons, and ligaments. Here we will review our recent work which has applied PSHG to study individual collagen fibrils, including investigation of how out of plane orientation affects PSHG images of collagen, and how differences in hydration alter collagen ultrastructure. We also present new data on the effect of applied tension on individual fibrils.

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Second harmonic generation microscopy of polysaccharide macrostructures

Cisek,

Tokarz

2024

Advanced Biophysical Techniques for Polysaccharides Characterization

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Investigation into the structure of crystalline maltodextrin particles by second harmonic generation microscopy

Cited by 5 publications

References 30 publications

Effect of out of plane orientation on polarization second harmonic generation of single collagen fibrils

Effect of out of plane orientation on polarization second harmonic generation of single collagen fibrils

Determining the helical pitch and relative polarity of collagen using polarization-resolved SHG

Second harmonic generation microscopy of polysaccharide macrostructures

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