2D/3D cryo x-ray fluorescence imaging at the bionanoprobe at the advanced photon source

Chen, S.; Paunesku, Tatjana; Yuan, Ye; Deng, Junjing; Jin, Qiaoling; Vine, D. J.; Lai, Barry; Hornberger, Benjamin; Brister, Keith; Jacobsen, Chris; Woloschak, Gayle E.; Vogt, Stefan

doi:10.1063/1.4937522

Cited by 5 publications

(4 citation statements)

References 16 publications

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“…The use of synchrotron-XRF for the analysis of metal anticancer complexes in vitro and ex vivo (e.g., in tissue from xenograft models) has been reported for platinum, − ruthenium, , iridium, and osmium − anticancer agents, among others. Notably, cryo-XRF is emerging at various beamlines for the analysis of frozen-hydrated cells or tissues close to their native state, including ID16A (ESRF, Grenoble) and 9-ID-B (APS, Illinois) …”

Section: Introductionmentioning

confidence: 99%

Single-Cell Chemistry of Photoactivatable Platinum Anticancer Complexes

et al. 2021

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The Pt(IV) prodrug trans, trans, trans-[Pt(pyridine) 2 (N 3 ) 2 (OH) 2 ] (Pt1) and its coumarin derivative trans, trans, trans-[Pt(pyridine) 2 (N 3 ) 2 (OH)(coumarin-3-carboxylate)] (Pt2) are promising agents for photoactivated chemotherapy. These complexes are inert in the dark but release Pt(II) species and radicals upon visible light irradiation, resulting in photocytotoxicity toward cancer cells. Here, we have used synchrotron techniques to investigate the in-cell behavior of these prodrugs and visualize, for the first time, changes in cellular morphology and Pt localization upon treatment with and without light irradiation. We show that photoactivation of Pt2 induces remarkable cellular damage with extreme alterations to multiple cellular components, including formation of vacuoles, while also significantly increasing the cellular accumulation of Pt species compared to dark conditions. X-ray absorption near-edge structure (XANES) measurements in cells treated with Pt2 indicate only partial reduction of the prodrug upon irradiation, highlighting that phototoxicity in cancer cells may involve not only Pt(II) photoproducts but also photoexcited Pt(IV) species.

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Section: Introductionmentioning

confidence: 99%

Single-Cell Chemistry of Photoactivatable Platinum Anticancer Complexes

et al. 2021

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“…13 The uptake of nanoparticles by cells was visualized using the Bionanoprobe at the APS. 14 An enhanced depth resolution can be achieved either by confocal XRF [15][16][17] or by 3D x-ray fluorescence tomography. [18][19][20] Particularly, for an analysis of single cells, x-ray fluorescence microscopy provides a useful addition to the toolbox of molecular biology as the elemental distribution inside ARTICLE avs.scitation.org/journal/bip cryogenically fixed cells, e.g., after treatment with drugs, [4][5][6]21,22 can be imaged without the use of labels.…”

Section: Introductionmentioning

confidence: 99%

Micro x-ray fluorescence analysis of trace element distribution in frozen hydrated HeLa cells at the P06 beamline at Petra III

et al. 2021

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X-ray fluorescence analysis enables the study of trace element distributions in biological specimens. When this analysis is done under cryogenic conditions, cells are cryofixed as closely as possible to their natural physiological state, and the corresponding intracellular elemental densities can be analyzed. Details about the experimental setup used for analysis at the P06 beamline at Petra III, DESY and the used cryo-transfer system are described in this work. The system was applied to analyze the elemental distribution in single HeLa cells, a cell line frequently used in a wide range of biological applications. Cells adhered to silicon nitride substrates were cryoprotected within an amorphous ice matrix. Using a continuous scanning scheme and a KB x-ray focus, the distribution of elements in the cells was studied. We were able to image the intracellular potassium and zinc levels in HeLa cells as two key elements relevant for the physiology of cells.

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“…All this complicates the measurements, since it requires specic equipment and technology, together with necessary expertise; however, it may produce exceptional results. Many examples in literature have shown the potential of cryogenic measurements for ptychography, 7,8 tomography, 9,10 XANES 11 and X-ray Fluorescence (XRF) spectro-imaging. 12 For these reasons, several synchrotron facilities have been equipped with appropriate sample environments.…”

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confidence: 99%

“…Yet, cryo-xation and sectioning of human or animal so tissues have remained a challenge. 7,8 Cryo-xation can be performed relatively easily and is widely used on small samples, such as cells, 6,[12][13][14][15] viruses, proteins etc., or porous specimens such as plants; especially for the latter, cryo-xation is currently the most popular sample preparation protocol. [16][17][18][19][20][21] A proper vitrication implies the formation of amorphous ice in the sample, which is crucial for its preservation, while an incomplete process would cause formation of the ice crystals leading to the breaking of membranes and distortion of the sample morphology.…”

mentioning

confidence: 99%