A drawer-type abdominal window with an acrylic/resin coverslip enables long-term intravital fluorescence/photoacoustic imaging of the liver

Deng, Deqiang; Dai, Bolei; Wei, Jianshuang; Yuan, Xuenan; Yang, Xiaoquan; Qi, Shuhong

doi:10.1515/nanoph-2021-0281

Cited by 13 publications

(14 citation statements)

References 40 publications

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“…This problem can be addressed by an implanted abdominal imaging window, which allows longitudinal IVM observation for up to 5 weeks and has been used for the visualization of metastasis formation in the liver over 14 days ( Ritsma et al, 2012 ). Deng et al further developed a drawer-type abdominal window to eliminate the impact of respiration and heartbeat on the quality of imaging ( Deng et al, 2021 ).…”

Section: Novel Ivm Methods To Investigate Dynamic Single-cell Behaviormentioning

confidence: 99%

Intravital Imaging of Inflammatory Response in Liver Disease

Wang

Wang²

2022

Front. Cell Dev. Biol.

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The healthy liver requires a strictly controlled crosstalk between immune and nonimmune cells to maintain its function and homeostasis. A well-conditioned immune system can effectively recognize and clear noxious stimuli by a self-limited, small-scale inflammatory response. This regulated inflammatory process enables the liver to cope with daily microbial exposure and metabolic stress, which is beneficial for hepatic self-renewal and tissue remodeling. However, the failure to clear noxious stimuli or dysregulation of immune response can lead to uncontrolled liver inflammation, liver dysfunction, and severe liver disease. Numerous highly dynamic circulating immune cells and sessile resident immune and parenchymal cells interact and communicate with each other in an incredibly complex way to regulate the inflammatory response in both healthy and diseased liver. Intravital imaging is a powerful tool to visualize individual cells in vivo and has been widely used for dissecting the behavior and interactions between various cell types in the complex architecture of the liver. Here, we summarize some new findings obtained with the use of intravital imaging, which enhances our understanding of the complexity of immune cell behavior, cell–cell interaction, and spatial organization during the physiological and pathological liver inflammatory response.

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Section: Novel Ivm Methods To Investigate Dynamic Single-cell Behaviormentioning

confidence: 99%

Intravital Imaging of Inflammatory Response in Liver Disease

Wang

Wang²

2022

Front. Cell Dev. Biol.

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“…2.1.3 | Dual-mode fluorescence/ photoacoustic imaging based on a drawer-type abdominal window A drawer-type abdominal window with an acrylic/resin coverslip model (DAWarc) was developed by our group. This model not only solves the liver focal plane dither caused by breathing and heart beating through the drawer structure designed to physically fix the hepatic lobes, but also balances the penetration efficiency of light and sound signals with the acrylic/resin cover, making it compatible with long-term live fluorescence and photoacoustic imaging [26]. Using the DAWarc model, long-term intravital fluorescence/photoacoustic imaging was performed for several days to reveal the spatial location and movement behavior of invariant NKT (iNKT) cells, as well as the distribution and morphological information of KCs in the microenvironment of liver metastases [26].…”

Section: Window Model For Intravital Optical Imaging Of the Livermentioning

confidence: 99%

Advances in intravital imaging of liver immunity using optical microscopy and labeling methods

et al. 2023

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The use of optical microscopy and labeling methods in intravital imaging allows for direct tracking of cell behavior and dynamic changes at the molecular level in the physiological or pathological microenvironment of living animals, revealing the spatiotemporal information of individual cells in the immune response. The liver is an immunological organ that contains unique innate and adaptive immune cells, including Kupffer cells (KCs) and different types of T cells, and is involved in coordinating multiple immune responses in the body. Using intravital imaging to visualize the movement behaviors and functions of immune cells during the reaction processes of the liver under physiological and pathological conditions has shed new light on the understanding of liver immunity, which is of great significance for the diagnosis and treatment of liver diseases. This review introduces various window models and labeling methods for the liver in intravital optical imaging and describes how it provides movement behavior and functional information about different types of immune cells, such as KCs and T cells, in the liver. Additionally, we highlight recent advances in intravital optical imaging of liver diseases, such as nonalcoholic fatty liver disease, infections, and tumors. This review aims to be a useful resource for comprehending the developments and achievements in intravital imaging of the liver and uncovering spatiotemporal information of immune response in a living microenvironment.

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“…Before performing long-term intravital imaging of the liver, mice needed to be assembled with a drawer-type abdominal window (DAW) [31]. The mice that successfully assembled DAW were immobilized in custom-made boxes, exposing the imaging window.…”

Section: Long-term Intravital Imaging Of Liversmentioning

confidence: 99%

“…In another investigation, we performed intravital molecular imaging to reveal the restrained capacity of CTLs to kill tumor cells in the liver through dynamically monitoring the fluorescence resonance energy transfer (FRET) signals of caspase-3 and calcium sensors in tumor cells [30]. Additionally, for long-term intravital imaging of the liver, we developed a drawer-type abdominal window with an acrylic/resin coverslip named DAWarc, which we used to observe the colonization and growth of liver metastasis for up to 10 days [31]. We expect intravital imaging to represent a powerful tool for further evaluating the tfRFP-elicited anti-tumor mechanism, especially its stimulation of specific humoral immune responses in the liver.…”

Section: Introductionmentioning

confidence: 99%

Intravital molecular imaging reveals that ROS-caspase-3-GSDME-induced cell punching enhances humoral immunotherapy targeting intracellular tumor antigens

Dai¹,

Zhang²,

Qi³

et al. 2022

Theranostics

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Tumor antigens (TAs)-induced humoral immune responses or TAs-specific antibodies have great application prospects for tumor therapy. However, more than half of TAs are intracellular antigens (intra-Ags) that are hardly recognized by antibodies. It is worthy to develop immunotherapeutic strategies for targeting intra-Ags. Methods: We used the far-red fluorescent protein tfRFP as an intracellular antigen to immunize mice and generated a liver metastasis model by injecting tfRFP-expressing B16 melanoma cells (tfRFP-B16) via the spleen. Intravital molecular imaging and atomic force microscopy were performed to visualize the formation of tfRFP antigen-antibody complexes (also known as immune complexes) and punched holes in cell membranes. Results:The results showed that the tfRFP-elicited immune responses inhibited the metastasis of tfRFP-expressing melanoma cells in the liver. In the circulating tfRFP-B16 tumor cells, elevated reactive oxygen species (ROS) induced slight caspase-3 activation, a probable key factor in the cleavage of gasdermin E (GSDME) proteins and punching of holes in the tumor cell membrane. Increased tumor cell membrane permeability led to the release of intra-Ag tfRFP and binding with anti-tfRFP antibodies. The formation of tfRFP antigen-antibody complexes on the membranes of tfRFP-B16 cells activated complement components to form membrane attack complexes to further destroy the cell membrane. Neutrophils were rapidly recruited, and F4/80 + macrophages phagocytized the dying tumor cells. Conclusion:The process of circulating tumor cell elimination in the tfRFP-immunized mice was triggered through the ROS-caspase-3-GSDME pathway to form intra-Ag-antibody immune complexes, which were involved in the activation of the complement system, as well as the recruitment of neutrophils and F4/80 + macrophages. An intra-Ag-elicited humoral immune response is a potent strategy for eliminating liver metastasis, which is unaffected by the liver immune tolerogenic status.

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A drawer-type abdominal window with an acrylic/resin coverslip enables long-term intravital fluorescence/photoacoustic imaging of the liver

Cited by 13 publications

References 40 publications

Intravital Imaging of Inflammatory Response in Liver Disease

Intravital Imaging of Inflammatory Response in Liver Disease

Advances in intravital imaging of liver immunity using optical microscopy and labeling methods

Intravital molecular imaging reveals that ROS-caspase-3-GSDME-induced cell punching enhances humoral immunotherapy targeting intracellular tumor antigens

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