Synchrotron-based Transmission X-ray Microscopy (TXM) Observations of Fully Hydrated Blood Platelets and Their Activation Process

Yang, Nuri; Nho, Hyun Woo; Kalegowda, Yogesh; Kim, Jin Bae; Song, Jaewoo; Shin, Hyun‐Joon; Yoon, Tae Hyun

doi:10.5012/bkcs.2014.35.9.2625

Cited by 5 publications

(3 citation statements)

References 25 publications

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“…Synchrotron based X-ray transmission images were acquired at 500 eV (wavelength of 2.48 nm) with over 10 10 photons/s (0.1% BW) photon flux from the 10D beamline of the Pohang Light Source (PLS, Pohang, Republic of Korea) 32 . Detailed description and schematic illustration of the TXM setup used in this study were presented in recent publications 32 33 . The incoming X-ray was focused on the sample by a condenser zone plate (CZP, 2 mm diameter, 110 nm outermost zone width), and the transmitted X-ray was magnified with an objective zone plate (OZP, 84 μm diameter, 38 nm fine zone width).…”

Section: Experimental Methodsmentioning

confidence: 99%

Nanoscale characterization of local structures and defects in photonic crystals using synchrotron-based transmission soft X-ray microscopy

Nho

Kalegowda

Shin

et al. 2016

Sci Rep

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For the structural characterization of the polystyrene (PS)-based photonic crystals (PCs), fast and direct imaging capabilities of full field transmission X-ray microscopy (TXM) were demonstrated at soft X-ray energy. PS-based PCs were prepared on an O2-plasma treated Si3N4 window and their local structures and defects were investigated using this label-free TXM technique with an image acquisition speed of ~10 sec/frame and marginal radiation damage. Micro-domains of face-centered cubic (FCC (111)) and hexagonal close-packed (HCP (0001)) structures were dominantly found in PS-based PCs, while point and line defects, FCC (100), and 12-fold symmetry structures were also identified as minor components. Additionally, in situ observation capability for hydrated samples and 3D tomographic reconstruction of TXM images were also demonstrated. This soft X-ray full field TXM technique with faster image acquisition speed, in situ observation, and 3D tomography capability can be complementally used with the other X-ray microscopic techniques (i.e., scanning transmission X-ray microscopy, STXM) as well as conventional characterization methods (e.g., electron microscopic and optical/fluorescence microscopic techniques) for clearer structure identification of self-assembled PCs and better understanding of the relationship between their structures and resultant optical properties.

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Section: Experimental Methodsmentioning

confidence: 99%

Nanoscale characterization of local structures and defects in photonic crystals using synchrotron-based transmission soft X-ray microscopy

Nho

Kalegowda

Shin

et al. 2016

Sci Rep

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“…Recently, scanning transmission X-ray microscopy (STXM) has shown its capabilities and these may complement the current limitations of WM TEM by providing a better contrast mechanism and penetration depth with enough spatial resolution (Benzerara et al, 2004;Nelson et al, 2010;Kwon et al, 2014;Yang et al, 2014;Nho et al, 2016;Nho & Yoon, 2017). In particular, the contrast mechanism of STXM, which is based on the difference in X-ray absorption at the pre-and postedges, allows us to distinguish Ca-rich DGs, which may provide much clearer and more DG-specific contrast than the electron-density-based contrast of TEM.…”

Section: Introductionmentioning

confidence: 99%

Identification of Ca-rich dense granules in human platelets using scanning transmission X-ray microscopy

Trinh

Kwon

Gerelkhuu

et al. 2020

J Synchrotron Radiat

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Whole-mount (WM) platelet preparation followed by transmission electron microscopy (TEM) observation is the standard method currently used to assess dense granule (DG) deficiency (DGD). However, due to the electron-density-based contrast mechanism in TEM, other granules such as α-granules might cause false DG detection. Here, scanning transmission X-ray microscopy (STXM) was used to identify DGs and minimize false DG detection of human platelets. STXM image stacks of human platelets were collected at the calcium (Ca) L 2,3 absorption edge and then converted to optical density maps. Ca distribution maps, obtained by subtracting the optical density maps at the pre-edge region from those at the post-edge region, were used to identify DGs based on the Ca richness. DGs were successfully detected using this STXM method without false detection, based on Ca maps for four human platelets. Spectral analysis of granules in human platelets confirmed that DGs contain a richer Ca content than other granules. The Ca distribution maps facilitated more effective DG identification than TEM which might falsely detect DGs. Correct identification of DGs would be important to assess the status of platelets and DG-related diseases. Therefore, this STXM method is proposed as a promising approach for better DG identification and diagnosis, as a complementary tool to the current WM TEM approach.

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“…Additionally, because of the electron density-based contrast mechanism, other granules such as α-granules might cause false detection of DGs. Recently, scanning transmission X-ray microscopy (STXM) has shown its capabilities that may complement current limitations of WM TEM by providing better contrast mechanism and penetration depth with enough spatial resolution (11-16) . Particularly, the contrast mechanism of STXM, which is based on the differences in X-ray absorption at the pre- and post-edges, allows us to distinguish Ca-rich DGs, which may provide much clearer and more DG specific contrast than electron density-based contrast of TEM.…”

Section: Introductionmentioning

confidence: 99%

Identification of Ca-rich dense granules in human platelets using scanning transmission X-ray microscopy

Trinh

Kwon

Gerelkhuu

et al. 2019

Preprint

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12Whole mount (WM) platelet preparations followed by transmission electron microscopy (TEM) observation is the 13 standard method currently used to assess dense granule (DG) deficiency (DGD). However, due to electron densi-14 ty-based contrast mechanism in TEM, other granules such as α -granules might cause false DGs detection. Herein, 15 scanning transmission X-ray microscopy (STXM), was used to identify DGs and minimize false DGs detection of 16 human platelets. STXM image stacks of human platelets were collected at the calcium (Ca) L 2,3 absorption edge 17 and then converted to optical density maps. Ca distribution maps obtained by subtracting the optical density map 18 at pre-edge region from those obtained at post-edge region were used for identification of DGs based on richness 19 of Ca. Dense granules were successfully detected by using STXM method without false detection based on Ca 20 maps for 4 human platelets. Spectral analysis of granules in human platelets confirmed that DGs contained richer 21 Ca content than other granules. Image analysis of Ca maps provided quantitative parameters which would be use-22 ful for developing image-based DG diagnosis models. Therefore, we would like to propose STXM as a promising 23 approach for better DG identification and DGD diagnosis, as a complementary tool to the current WM TEM ap-24 proach. 25

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Synchrotron-based Transmission X-ray Microscopy (TXM) Observations of Fully Hydrated Blood Platelets and Their Activation Process

Cited by 5 publications

References 25 publications

Nanoscale characterization of local structures and defects in photonic crystals using synchrotron-based transmission soft X-ray microscopy

Nanoscale characterization of local structures and defects in photonic crystals using synchrotron-based transmission soft X-ray microscopy

Identification of Ca-rich dense granules in human platelets using scanning transmission X-ray microscopy

Identification of Ca-rich dense granules in human platelets using scanning transmission X-ray microscopy

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