To visualize scaffolds by confocal microscopy, we conjugated Hilyte Fluor 647 (Anaspec) to alginate using standard EDC/NHS chemistry, then mixed 1 part of the conjugate with 9 parts of GFOGER-modified alginate to fabricate scaffolds as described above. T cells were labeled with CellTracker Orange CMTMR (Invitrogen) immediately before seeding into these scaffolds. Three days post-implantation, the tumor resection bed was snap-frozen in OCT (Tissue-Tek) to produce cryosections, which were fixed with 2% paraformaldehyde, mounted in ProLong Gold Antifade reagent (Invitrogen), and imaged with a Zeiss LSM 780 NLO laser scanning confocal microscope.
Lsm 780 nlo laser
Manufactured by Zeiss
The LSM 780 NLO is a laser scanning microscope designed for advanced imaging applications. It features a tunable two-photon excitation laser that enables deep tissue imaging and fluorescence detection. The system provides high-resolution, non-invasive imaging capabilities for a wide range of samples.
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2 protocols using lsm 780 nlo laser
1
Visualizing Scaffold-Seeded T Cells
2
Visualizing Scaffold-Seeded T Cells
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To visualize scaffolds by confocal microscopy, we conjugated Hilyte Fluor 647 (Anaspec) to alginate using standard EDC/NHS chemistry, then mixed 1 part of the conjugate with 9 parts of GFOGER-modified alginate to fabricate scaffolds as described above. T cells were labeled with CellTracker Orange CMTMR (Invitrogen) immediately before seeding into these scaffolds. Three days post-implantation, the tumor resection bed was snap-frozen in OCT (Tissue-Tek) to produce cryosections, which were fixed with 2% paraformaldehyde, mounted in ProLong Gold Antifade reagent (Invitrogen), and imaged with a Zeiss LSM 780 NLO laser scanning confocal microscope.
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