In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish

Miskolci, Veronika; Tweed, Kelsey; Lasarev, Michael; Britt, Emily C; Walsh, Alex J.; Zimmerman, Landon J; McDougal, Courtney E; Cronan, Mark R.; Fan, Jing; Sauer, John-Demian; Skala, Melissa C.; Huttenlocher, Anna

doi:10.7554/elife.66080

Cited by 32 publications

(44 citation statements)

References 56 publications

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“…OMI has also been used to classify subsets of macrophages in monoculture, tumor coculture, and in vivo in zebrafish, and to distinguish between categories of blood cells ( i.e . erythrocytes, monocytes, granulocytes, lymphocytes) ( 42–44 ). Prior work also shown that NADH autofluorescence intensity increases in activated B cells compared to unstimulated B cells ( 45 ).…”

Section: Introductionmentioning

confidence: 99%

Autofluorescence lifetime imaging classifies human lymphocyte activation and subtype

Schmitz

Tweed

Rehani

et al. 2023

Preprint

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New non-destructive tools are needed to reliably assess lymphocyte function for immune profiling and adoptive cell therapy. Optical metabolic imaging (OMI) is a label-free method that measures the autofluorescence intensity and lifetime of metabolic cofactors NAD(P)H and FAD to quantify metabolism at a single-cell level. Here, we investigate whether OMI can resolve metabolic changes between human quiescent versus IL4/CD40 activated B cells and IL12/IL15/IL18 activated memory-like NK cells. We found that quiescent B and NK cells were more oxidized compared to activated cells. Additionally, the NAD(P)H mean fluorescence lifetime decreased and the fraction of unbound NAD(P)H increased in the activated B and NK cells compared to quiescent cells. Machine learning classified B cells and NK cells according to activation state (CD69+) based on OMI parameters with up to 93.4% and 92.6% accuracy, respectively. Leveraging our previously published OMI data from activated and quiescent T cells, we found that the NAD(P)H mean fluorescence lifetime increased in NK cells compared to T cells, and further increased in B cells compared to NK cells. Random forest models based on OMI classified lymphocytes according to subtype (B, NK, T cell) with 97.8% accuracy, and according to activation state (quiescent or activated) and subtype (B, NK, T cell) with 90.0% accuracy. Our results show that autofluorescence lifetime imaging can accurately assess lymphocyte activation and subtype in a label-free, non-destructive manner.

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Section: Introductionmentioning

confidence: 99%

Autofluorescence lifetime imaging classifies human lymphocyte activation and subtype

Schmitz

Tweed

Rehani

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…There are only a few works with optical metabolic imaging of the immune cells so far. For example, in 2020–2022 the group by M. Skala reported on in vitro [ 18 ] and intravital [ 26 , 27 ] studies of macrophages metabolism in normal and cancerous tissue using the intensity-based redox ratio and FLIM of NAD(P)H and FAD. Moreover, in 2021, they demonstrated the ability of autofluorescence imaging to distinguish between quiescent and activated T-cells, as well as between the subsets of CD4+ and CD8+ cells [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

FLIM of NAD(P)H in Lymphatic Nodes Resolves T-Cell Immune Response to the Tumor

Izosimova

Shirmanova

Shcheslavskiy

et al. 2022

IJMS

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Assessment of T-cell response to the tumor is important for diagnosis of the disease and monitoring of therapeutic efficacy. For this, new non-destructive label-free methods are required. Fluorescence lifetime imaging (FLIM) of metabolic coenzymes is a promising innovative technology for the assessment of the functional status of cells. The purpose of this work was to test whether FLIM can resolve metabolic alterations that accompany T-cell reactivation to the tumors. The study was carried out on C57Bl/6 FoxP3-EGFP mice bearing B16F0 melanoma. Autofluorescence of the immune cells in fresh lymphatic nodes (LNs) was investigated. It was found that fluorescence lifetime parameters of nicotinamide adenine dinucleotide (phosphate) NAD(P)H are sensitive to tumor development. Effector T-cells in the LNs displayed higher contribution of free NADH, the form associated with glycolysis, in all tumors and the presence of protein-bound NADPH, associated with biosynthetic processes, in the tumors of large size. Flow cytometry showed that the changes in the NADH fraction of the effector T-cells correlated with their activation, while changes in NADPH correlated with cell proliferation. In conclusion, FLIM of NAD(P)H in fresh lymphoid tissue is a powerful tool for assessing the immune response to tumor development.

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“…Thus, zebrafish are more helpful in experimental replication. (3) Zebrafish embryos and larvae are transparent, which allows for real-time observations of fluorescently labeled activities inside zebrafish. , On the contrary, mouse embryos are not transparent and they naturally develop inside the mother, so currently, it is impossible to observe live embryo development or any fluorescently labeled activities in the body. Nonetheless, uses of zebrafish as an in vivo model have received many doubts and concerns, and a key question is whether the BBB of zebrafish is qualified to mimic human’s.…”

Section: Carbon Dots In Nanomedicinementioning

confidence: 99%

Structure–Property–Activity Relationships in Carbon Dots

2022

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Carbon dots (CDs) are one of the most versatile nanomaterials discovered in the 21st century. They possess many properties and thus hold potentials in diverse applications. While an increasing amount of attention has been given to these novel nanoparticles, the broad scientific community is actively engaged in exploring their limits. Recent studies on the fractionalization and assembly of CDs further push the limits beyond just CDs and demonstrate that CDs are both a mixture of heterogeneous fractions and promising building blocks for assembly of large carbon-based materials. With CDs moving forward toward both microscopic and macroscopic levels, a good understanding of the structure−property−activity relationships is essential to forecasting the future of CDs. Hence, in this Perspective, structure−property− activity relationships are highlighted based on the repeatedly verified findings in CDs. In addition, studies on CD fractionalization and assembly are briefly summarized in this Perspective. Eventually, these structure−property−activity relationships and controllability are essential for the development of CDs with desired properties for various applications especially in photochemistry, electrochemistry, nanomedicine, and surface chemistry. In summary, in our opinion, since 2004 until the present, history has witnessed a great development of CDs although there is still some room for more studies. Also, considering many attractive properties, structure−property−activity relationships, and the building block nature of CDs, a variety of carbon-based materials of interest can be constructed from CDs with control. They can help reduce blind trials in the development of carbon-based materials, which is of great significance in materials science, chemistry, and any fields related to the applications.

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In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish

Cited by 32 publications

References 56 publications

Autofluorescence lifetime imaging classifies human lymphocyte activation and subtype

Autofluorescence lifetime imaging classifies human lymphocyte activation and subtype

FLIM of NAD(P)H in Lymphatic Nodes Resolves T-Cell Immune Response to the Tumor

Structure–Property–Activity Relationships in Carbon Dots

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