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(A) Schematic of the inverted widefield frequency-domain <t>FLIM</t> system used for live oxygen measurements. The setup includes a 470-nm high-power LED under external modulation, an inverted fluorescence microscope with appropriate excitation/emission filters (Ex 420–490 nm/Em 570–670 nm), and <t>a</t> <t>dual-tap</t> FLIM camera for multi-phase acquisition. (B) Confocal image of a cerebral organoid after 26 days in culture, stained with CellMask (green) to delineate tissue architecture and to localize the oxygen-sensitive CPOx microbeads (red). Scale bar: 100 μm.
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(A) Schematic of the inverted widefield frequency-domain <t>FLIM</t> system used for live oxygen measurements. The setup includes a 470-nm high-power LED under external modulation, an inverted fluorescence microscope with appropriate excitation/emission filters (Ex 420–490 nm/Em 570–670 nm), and <t>a</t> <t>dual-tap</t> FLIM camera for multi-phase acquisition. (B) Confocal image of a cerebral organoid after 26 days in culture, stained with CellMask (green) to delineate tissue architecture and to localize the oxygen-sensitive CPOx microbeads (red). Scale bar: 100 μm.
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(A) Schematic of the inverted widefield frequency-domain FLIM system used for live oxygen measurements. The setup includes a 470-nm high-power LED under external modulation, an inverted fluorescence microscope with appropriate excitation/emission filters (Ex 420–490 nm/Em 570–670 nm), and a dual-tap FLIM camera for multi-phase acquisition. (B) Confocal image of a cerebral organoid after 26 days in culture, stained with CellMask (green) to delineate tissue architecture and to localize the oxygen-sensitive CPOx microbeads (red). Scale bar: 100 μm.

Journal: Bio-protocol

Article Title: Characterizing Tissue Oxygen Tension During Neurogenesis in Human Cerebral Organoids

doi: 10.21769/BioProtoc.5507

Figure Lengend Snippet: (A) Schematic of the inverted widefield frequency-domain FLIM system used for live oxygen measurements. The setup includes a 470-nm high-power LED under external modulation, an inverted fluorescence microscope with appropriate excitation/emission filters (Ex 420–490 nm/Em 570–670 nm), and a dual-tap FLIM camera for multi-phase acquisition. (B) Confocal image of a cerebral organoid after 26 days in culture, stained with CellMask (green) to delineate tissue architecture and to localize the oxygen-sensitive CPOx microbeads (red). Scale bar: 100 μm.

Article Snippet: Dual-tap complementary metal-oxide semiconductor (CMOS) FLIM camera (Excelitas Technologies Corp., catalog number: PCO.FLIM) 5.

Techniques: Fluorescence, Microscopy, Staining