Confocal Real-Time Analysis of Cutaneous Platelet Recruitment during Immune Complex‒Mediated Inflammation

Currie, S.; Stegmeyer, Rebekka I; Mildner, Karina; Breitsprecher, Leonhard; Zeuschner, Dagmar; Psathaki, Olympia E.; Schäfer, Kerstin; Wilkens, Markus; Volkery, Stefan; Nieswandt, Bernhard; Vestweber, Dietmar

doi:10.1016/j.jid.2022.03.011

Cited by 4 publications

(10 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A titanium dorsal skinfold chamber frame (small dorsal kit, APJ trading Co Inc) was surgically attached to the disinfected skin, and a circular window area (~ 12 mm diameter) was prepared for microscopy. For further details, see Currie et al (2022).…”

Section: Dorsal Skinfold Chamber Ivmmentioning

confidence: 99%

“…2a, b). Inflammation was induced by initiating the reverse passive Arthus reaction, as described by Currie et al (2022). Prior to anesthesia, mice were i.v.…”

Section: Dorsal Skinfold Chamber Ivmmentioning

confidence: 99%

“…SBF-SEM allows the visualization of almost unlimited 3D volumes, capturing complex cell-cell interactions. This turned out to be the fundamental instrument to solve the original biological question, as platelets could be detected in full size transferring the basement membrane barrier of the activated vessel (as documented in detail in Currie et al 2022).…”

Section: Outlook and Conclusionmentioning

confidence: 99%

“…The question at hand could only be answered sufficiently by three-dimensional volume electron microscopy, here using serial blockface scanning electron microscopy (SBF-SEM) (Denk and Horstmann 2004). As a consequence, extra adjustments for the sample preparation were incorporated into the workflow (Currie et al 2022).…”

Section: Introductionmentioning

confidence: 99%

“…As a consequence, extra adjustments for the sample preparation were incorporated into the workflow (Currie et al. 2022 ).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Landmark-based retrieval of inflamed skin vessels enabled by 3D correlative intravital light and volume electron microscopy

Mildner

Breitsprecher

Currie

et al. 2022

Histochem Cell Biol

Self Cite

View full text Add to dashboard Cite

The nanometer spatial resolution of electron microscopy imaging remains an advantage over light microscopy, but the restricted field of view that can be inspected and the inability to visualize dynamic cellular events are definitely drawbacks of standard transmission electron microscopy (TEM). Several methods have been developed to overcome these limitations, mainly by correlating the light microscopical image to the electron microscope with correlative light and electron microscopy (CLEM) techniques. Since there is more than one method to obtain the region of interest (ROI), the workflow must be adjusted according to the research question and biological material addressed. Here, we describe in detail the development of a three-dimensional CLEM workflow for mouse skin tissue exposed to an inflammation stimulus and imaged by intravital microscopy (IVM) before fixation. Our aim is to relocate a distinct vessel in the electron microscope, addressing a complex biological question: how do cells interact with each other and the surrounding environment at the ultrastructural level? Retracing the area over several preparation steps did not involve any specific automated instruments but was entirely led by anatomical and artificially introduced landmarks, including blood vessel architecture and carbon-coated grids. Successful retrieval of the ROI by electron microscopy depended on particularly high precision during sample manipulation and extensive documentation. Further modification of the TEM sample preparation protocol for mouse skin tissue even rendered the specimen suitable for serial block-face scanning electron microscopy (SBF-SEM).

show abstract

Section: Dorsal Skinfold Chamber Ivmmentioning

confidence: 99%

“…2a, b). Inflammation was induced by initiating the reverse passive Arthus reaction, as described by Currie et al (2022). Prior to anesthesia, mice were i.v.…”

Section: Dorsal Skinfold Chamber Ivmmentioning

confidence: 99%

Section: Outlook and Conclusionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…As a consequence, extra adjustments for the sample preparation were incorporated into the workflow (Currie et al. 2022 ).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Landmark-based retrieval of inflamed skin vessels enabled by 3D correlative intravital light and volume electron microscopy

Mildner

Breitsprecher

Currie

et al. 2022

Histochem Cell Biol

Self Cite

View full text Add to dashboard Cite

show abstract

Platelet Plugs Prevent Vascular Hemorrhage at Sites of Neutrophil Diapedesis

Vargas

Simon

2022

Journal of Investigative Dermatology

View full text Add to dashboard Cite

The localization, origin, and impact of platelets in the tumor microenvironment are tumor type-dependent

Le Chapelain,

Jadoui,

Gros

et al. 2024

J Exp Clin Cancer Res

Self Cite

View full text Add to dashboard Cite

Background How platelets interact with and influence the tumor microenvironment (TME) remains poorly characterized. Methods We compared the presence and participation of platelets in the TME of two tumors characterized by highly different TME, PyMT AT-3 mammary tumors and B16F1 melanoma. Results We show that whereas firmly adherent platelets continuously line tumor vessels of both AT-3 and B16F1 tumors, abundant extravascular stromal clusters of platelets from thrombopoietin-independent origin were present only in AT-3 mammary tumors. We further show that platelets influence the angiogenic and inflammatory profiles of AT-3 and B16F1 tumors, though with very different outcomes according to tumor type. Whereas thrombocytopenia increased bleeding in both tumor types, it further caused severe endothelial degeneration associated with massive vascular leakage, tumor swelling, and increased infiltration of cytotoxic cells, only in AT-3 tumors. Conclusions These results indicate that while platelets are integral components of solid tumors, their localization and origin in the TME, as well as their impact on its shaping, are tumor type-dependent.

show abstract

Confocal Real-Time Analysis of Cutaneous Platelet Recruitment during Immune Complex‒Mediated Inflammation

Cited by 4 publications

References 34 publications

Landmark-based retrieval of inflamed skin vessels enabled by 3D correlative intravital light and volume electron microscopy

Landmark-based retrieval of inflamed skin vessels enabled by 3D correlative intravital light and volume electron microscopy

Platelet Plugs Prevent Vascular Hemorrhage at Sites of Neutrophil Diapedesis

The localization, origin, and impact of platelets in the tumor microenvironment are tumor type-dependent

Contact Info

Product

Resources

About