Structural and elemental changes in glioblastoma cells in situ: complementary imaging with high resolution visible light- and X-ray microscopy

Abstract

The glioblastoma (GBM) is characterized by a short median survival and an almost 100% tumor related mortality. GBM cells exhibit highly invasive behavior whose mechanisms are not yet fully understood. The present study explores application of X-ray and visible light microscopy to display the elemental and structural images of cells from 3 patient derived GMB samples and an established GMB cell line. Slight differences in elemental concentrations, in actin cytoskeleton organization and cell morphology were noted between all cells types by X-ray fluorescence and full field soft X-ray microscopy, as well as the Structured Illumination Super-resolution Microscope (SIM). Different sample preparation approaches were used to match each imaging technique. While preparation for SIM included cell fixation and staining, intact frozen hydrated cells were used for the trace element imaging by hard X-ray fluorescence and exploration of the structural features by soft X-ray absorption tomography. Each technique documented differences between samples with regard to morphology and elemental composition and underscored the importance of use of multiple patient derived samples for detailed GBM study.

PLEK2, RRM2, GCSH: A Novel WWOX-Dependent Biomarker Triad of Glioblastoma at the Crossroads of Cytoskeleton Reorganization and Metabolism Alterations

“…GBM may develop de novo (the primary tumor) or via malignant progression from lower-grade glioma (LGG) as glioblastoma IDH mutated [3]-regardless of derivation, patient's prognosis is dismal [4]. The malignant phenotype of GBM results from, e.g., dynamics of cytoskeleton [5,6], which has been found to guide signaling pathways [7], proliferation [8], polarity [9], cell cycle [10], or epithelial-to-mesenchymal transition (EMT) [11] and metastasis [12]. Ultimately, cytoskeleton controls two processes that impact on cancer malignant behavior, i.e., cellular division and invasion [6].…”

Lipids status and copper in a single astrocyte of the rat model for amyotrophic lateral sclerosis: Correlative synchrotron‐based X‐ray and infrared imaging

“…With the aim to obtain information regarding the subcellular distribution of redox-active trace elements Cu and Fe, as well as Zn, we employed the XRF microscopy at the APS synchrotron facility. The XRF microscope installed on the Bionanoprobe bio-imaging stations provided the sample orientation necessary for this correlative imaging and high spatial and chemical resolution for hard X-ray imaging of single cells [34]. Moreover, the online visible light microscope provided better precision than a micrometer for the positioning of investigated cells.…”

Multifunctionalized Carbon Dots as an Active Nanocarrier for Drug Delivery to the Glioblastoma Cell Line

“…29,36,37 Besides, we showed that elemental composition and cytoskeletal structure differ in the same patient-derived GBM cells, as studied by synchrotron radiation-based soft-X-ray tomography and hard X-ray fluorescence. 35 Thus, as a step forward, in this study, we first synthesized characterized and afterward tested a system based on carbon nanoparticles from the 2-acrylamido-2-methylpropanesulfonic acid (AMPS), as a possible organic delivery system (AMPS-CDs NPs), for the drug riluzole in GBM cells. We chose AMPS as a potential nanocarrier, as it possesses free sulphonyl, carboxyl, and amide groups.…”

“…Besides, riluzole directly affects biomacromolecule changes, namely, carbohydrate metabolism, DNA structure, lipid structures, and protein secondary conformation. 28 Riluzole is approved by the Food and Drug Administration (FDA) as an amyotrophic lateral sclerosis (ALS) drug, 29 and recent studies showed that treatment with riluzole suppresses the proliferation of many cancer cells 30 − 35 and GBM. 29 , 36 , 37 Besides, we showed that elemental composition and cytoskeletal structure differ in the same patient-derived GBM cells, as studied by synchrotron radiation-based soft-X-ray tomography and hard X-ray fluorescence.…”

“… 29 , 36 , 37 Besides, we showed that elemental composition and cytoskeletal structure differ in the same patient-derived GBM cells, as studied by synchrotron radiation-based soft-X-ray tomography and hard X-ray fluorescence. 35 …”