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Inserting ‘OFF-to-ON’ BODIPY tags into cytokines: a fluorogenic interleukin IL33 for real-time imaging of immune cells

ACS Cent. Sci. 2023, doi: 10.1021/acscentsci.3c01125

Time-lapse fluorescence confocal microscopy images of transfected HEK-Blue cells after incubation with IL-33(6) (green) and nuclear counterstain DRAQ5 (red). IL-33(6) was added as fluorescence images were recorded. Excitation lasers: 488 nm (for IL-33(6)), 561 nm (for DRAQ5). Movie recorded for 5 min and compressed (jpeg) at 10 fps.

Fluorogenic granzyme A substrates enable real-time imaging of adaptive immune cell activity

Angew. Chem. Int. Ed. 2023, e202216142, doi: 10.1002/anie.202216142

Time-lapse fluorescence microscopy of live co-cultures of CD8+ T cells (counterstained with CellTrackerTM Orange, red) and EL4 cancer cells. Cells were incubated with compound 10 (3 µM, magenta for GzmA activity) and probe H5 (2.5 µM, green for GzmB activity). Movie recorded at 1 frame every 2 minutes.

A fluorogenic probe for granzyme B enables in-biopsy evaluation and screening of response to anticancer immunotherapies

Nat. Commun. 2022, 13, 2366, doi: 10.1038/s41467-022-29691-w

Real-time imaging of probe H5 in co-cultures of OT-I CD8+ T cells and OVA-EL4 cancer cells. Time-course widefield microscope images of OT-I CD8+ T cells (counterstained with Cell Tracker Orange, red) killing OVA-EL4 cancer cells in the presence of probe H5 (10 µM, green) and the dead marker Sytox Blue (1 µM, blue). Scale bar: 10 µm.

A bivalent activatable fluorescent probe for screening and intravital imaging of chemotherapy-induced cancer cell death

Angew. Chem. Int. Ed. 2022, doi: 10.1002/anie.202113020

Time-course intravital images of cisplatin-treated mammary tumours highlighting ECFP+ cancer cells (blue), EGFP+ macrophages (green) and apoptotic cells after in vivo administration of Apotracker Red (red). A recruited macrophage moves towards apoptotic tumour cells, whereas the white circles feature a macrophage in close proximity to Apotracker Red-stained tumour cells. Scale bar: 30 µm.

Photoactivatable metabolic warheads enable precise and safe ablation of target cells in vivo

Nat. Commun. 2021, doi: 10.1038/s41467-021-22578-2

LEFT VIDEO: Recording of swimming motion of a representative 3 dpf zebrafish larva after treatment with SeNBD-glucose (compound 15) and light irradiation. Larvae were placed in a 10‐cm Petri dish filled with Danieau’s solution.

RIGHT VIDEO:
 Heartbeat monitoring of a representative 3 dpf zebrafish larva after treatment with SeNBD-glucose (compound 15) and light irradiation. Larvae were anesthetized with MS222 and mounted sideways in 3% methylcellulose for continuous heartbeat recording using a Hamamatsu camera (40 frames per second).

A palette of minimally-tagged sucrose analogues for real-time Raman imaging of intracellular plant metabolism

Angew. Chem. Int. Ed. 2021doi: 10.1002/anie.202016802

LEFT VIDEO: Recording of BY2 cells after co-incubation of sucrose and alkyne-sucrose 6. Longitudinal Raman imaging was acquired at 2,116 cm-1 (10 frames per second).

 

RIGHT VIDEO: Recording of BY2 cells after incubation with alkyne-sucrose 6 alone. Longitudinal Raman imaging was acquired at 2,116 cm-1 (10 frames per second).

A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis.

Nat. Commun. 2020doi: 10.1038/s41467-020-17772-7

LEFT VIDEO: Early-stage imaging of BL-2 cells undergoing apoptosis with Apo-15. Human BL-2 cells were UV-irradiated (300 mJ cm-2) and incubated for 3 h at 37°C and 5% CO2. Time-lapse wash-free imaging was performed immediately after addition of Apo-15 (100 nM) using a spinning-disk microscope (Andor) under live-cell conditions (exc.: 488 nm, em.: 525 nm). Images were acquired every 30 s for up to 15 min. Scale bar 10 µm.

 

RIGHT VIDEO: Apo-15 labels subcellular material from apoptotic neutrophils. Neutrophils were isolated from the peripheral blood of healthy volunteers with Percoll density gradient and treated with R-roscovitine (20 µM) prior to imaging. Time-lapse wash-free imaging was performed immediately after addition of Apo-15 (100 nM) using a spinning-disk microscope (Andor) under live-cell conditions (exc.: 488 nm, em.: 525 nm). Images were acquired every 1 s for up to 10 min. Scale bar: 10 µm.

LEFT VIDEO: Time-lapse TIRF microscopy movie corresponding to multiple particle tracking in untreated normoxic HeLa cells after incubation with compound 10 (100 uM). Time is indicated as seconds and images were acquired every 50 ms and played at 30 frames per second. Scale bar: 5 µm.

 

RIGHT VIDEO: Time-lapse TIRF microscopy movie corresponding to multiple particle tracking in DMOG-treated hypoxic HeLa cells  after incubation with compound 10 (100 uM). Time is indicated as seconds and images were acquired every 50 ms and played at 30 frames per second. Scale bar: 5 µm.

Chemical modulation of in vivo macrophage function with subpopulation-specific fluorescent prodrug conjugates.

ACS Cent. Sci., 2017, doi:10.1021/acscentsci.7b00262

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LEFT VIDEO: Time-lapse video showing mCherry-expressing macrophages at the wound edge at 28 h post wounding and 24 h post compound 5 administration. Several mCherry-expressing macrophages contain fluorescent phagosomes (yellow arrows) and they show  rounded morphology and immobile phenotype, suggesting that these cells are undergoing cell death. Scale bar: 50 μm.

RIGHT VIDEO: Time-lapse video showing mCherry-expressing macrophages at the wound edge within 4 h post wounding and 30 min of compound 5 administration. White arrows point at only few macrophages containing fluorescent phagosomes. Macrophages are still actively engulfing and moving, suggesting that they are still viable at this stage. Scale bar: 50 μm

Spacer-free BODIPY fluorogens in antimicrobial peptides for direct imaging of fungal infection in human tissue.

Nat. Commun., 2016, doi: 10.1038/ncomms10940.