Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy

Kim, Hong-Lim; Riew, Tae-Ryong; Park, Jieun; Lee, Youngchun; Kim, In‐Beom

doi:10.3390/ijms22084273

Cited by 5 publications

(4 citation statements)

References 23 publications

(15 reference statements)

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“…This technique is a better choice to preserve the ultrastructure. Recent developments exist that combine the benefits of both techniques ( Jones 2016 ; Kim et al, 2021 ).…”

Section: Electron and X-ray Based Microscopy Techniquesmentioning

confidence: 99%

Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale

Vanslembrouck

Chen

Larabell

et al. 2022

Front. Cell Dev. Biol.

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Considerable progress has been made in our knowledge of the morphological and functional varieties of anchoring junctions. Cell-cell adhesion contacts consist of discrete junctional structures responsible for the mechanical coupling of cytoskeletons and allow the transmission of mechanical signals across the cell collective. The three main adhesion complexes are adherens junctions, tight junctions, and desmosomes. Microscopy has played a fundamental role in understanding these adhesion complexes on different levels in both physiological and pathological conditions. In this review, we discuss the main light and electron microscopy techniques used to unravel the structure and composition of the three cell-cell contacts in epithelial and endothelial cells. It functions as a guide to pick the appropriate imaging technique(s) for the adhesion complexes of interest. We also point out the latest techniques that have emerged. At the end, we discuss the problems investigators encounter during their cell-cell adhesion research using microscopic techniques.

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“…This technique is a better choice to preserve the ultrastructure. Recent developments exist that combine the benefits of both techniques ( Jones 2016 ; Kim et al, 2021 ).…”

Section: Electron and X-ray Based Microscopy Techniquesmentioning

confidence: 99%

Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale

Vanslembrouck

Chen

Larabell

et al. 2022

Front. Cell Dev. Biol.

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“…Sections were labeled with DAPI for 10 min. Immunolabeled sections were photographed using confocal microscopy and processed for electron microscopy as previously described (Riew et al, 2019(Riew et al, , 2022Kim et al, 2021).…”

Section: Electron Microscopic Analysismentioning

confidence: 99%

Astrocytes are involved in the formation of corpora amylacea-like structures from neuronal debris in the CA1 region of the rat hippocampus after ischemia

Riew,

Hwang,

Jin

et al. 2023

Front. Cell. Neurosci.

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Recently, we demonstrated that the corpora amylacea (CA), a glycoprotein-rich aggregate frequently found in aged brains, accumulates in the ischemic hippocampus and that osteopontin (OPN) mediates the entire process of CA formation. Therefore, this study aimed to elucidate the mechanisms by which astrocytes and microglia participate in CA formation during the late phase (4–12 weeks) of brain ischemia. Based on various morphological analyses, including immunohistochemistry, in situ hybridization, immunoelectron microscopy, and correlative light and electron microscopy, we propose that astrocytes are the primary cells responsible for CA formation after ischemia. During the subacute phase after ischemia, astrocytes, rather than microglia, express Opn messenger ribonucleic acid and OPN protein, a surrogate marker and key component of CA. Furthermore, the specific localization of OPN in the Golgi complex suggests that it is synthesized and secreted by astrocytes. Astrocytes were in close proximity to type I OPN deposits, which accumulated in the mitochondria of degenerating neurons before fully forming the CA (type III OPN deposits). Throughout CA formation, astrocytes remained closely attached to OPN deposits, with their processes exhibiting well-developed gap junctions. Astrocytic cytoplasmic protein S100β, a calcium-binding protein, was detected within the fully formed CA. Additionally, ultrastructural analysis revealed direct contact between astroglial fibrils and the forming facets of the CA. Overall, we demonstrated that astrocytes play a central role in mediating CA formation from the initial stages of OPN deposit accumulation to the evolution of fully formed CA following transient ischemia in the hippocampus.

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“…Coverslipped sections were examined with a confocal microscope and photographed at ×200, ×400, and × 630 magnifications with a differential interference contrast setting to identify specific areas for succeeding electron microscopy. After the sections were removed from the coverslips, the tissues were prepared for electron microscopy as described previously (Kim et al, 2021;Riew et al, 2021).…”

Section: Immunoelectron Microscopymentioning

confidence: 99%

Infiltration of meningeal macrophages into the Virchow–Robin space after ischemic stroke in rats: Correlation with activated PDGFR-β-positive adventitial fibroblasts

Riew

Hwang

Jin

et al. 2022

Front. Mol. Neurosci.

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Macrophages play a crucial role in wound healing and fibrosis progression after brain injury. However, a detailed analysis of their initial infiltration and interaction with fibroblasts is yet to be conducted. This study aimed to investigate the possible route for migration of meningeal macrophages into the ischemic brain and whether these macrophages closely interact with neighboring platelet-derived growth factor beta receptor (PDGFR-β)-positive adventitial fibroblasts during this process. A rat model of ischemic stroke induced by middle cerebral artery occlusion (MCAO) was developed. In sham-operated rats, CD206-positive meningeal macrophages were confined to the leptomeninges and the perivascular spaces, and they were not found in the cortical parenchyma. In MCAO rats, the number of CD206-positive meningeal macrophages increased both at the leptomeninges and along the vessels penetrating the cortex 1 day after reperfusion and increased progressively in the extravascular area of the cortical parenchyma by 3 days. Immunoelectron microscopy and correlative light and electron microscopy showed that in the ischemic brain, macrophages were frequently located in the Virchow–Robin space around the penetrating arterioles and ascending venules at the pial surface. This was identified by cells expressing PDGFR-β, a novel biomarker of leptomeningeal cells. Macrophages within penetrating vessels were localized in the perivascular space between smooth muscle cells and PDGFR-β-positive adventitial fibroblasts. In addition, these PDGFR-β-positive fibroblasts showed morphological and molecular characteristics similar to those of leptomeningeal cells: they had large euchromatic nuclei with prominent nucleoli and well-developed rough endoplasmic reticulum; expressed nestin, vimentin, and type I collagen; and were frequently surrounded by collagen fibrils, indicating active collagen synthesis. In conclusion, the perivascular Virchow–Robin space surrounding the penetrating vessels could be an entry route of meningeal macrophages from the subarachnoid space into the ischemic cortical parenchyma, implying that activated PDGFR-β-positive adventitial fibroblasts could be involved in this process.

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Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy

Cited by 5 publications

References 23 publications

Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale

Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale

Astrocytes are involved in the formation of corpora amylacea-like structures from neuronal debris in the CA1 region of the rat hippocampus after ischemia

Infiltration of meningeal macrophages into the Virchow–Robin space after ischemic stroke in rats: Correlation with activated PDGFR-β-positive adventitial fibroblasts

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