Primary keratinocytes were isolated from WT and Twist1 KO mice as previously described (35 ), plated evenly on collagen-coated 6-well culture plates, and grown until confluent. Cells were wounded by scratching longitudinally with a 200 μl pipette tip and incubated in complete medium for 24 hours. Cells were imaged with a Nikon Ti-E inverted light microscope.
Ti e inverted light microscope
Manufactured by Nikon
The Nikon Ti-E inverted light microscope is a versatile research-grade instrument designed for a wide range of microscopy applications. It features a stable, inverted design that allows for the observation of samples from below. The Ti-E provides excellent optical performance and is equipped with a range of advanced features to support various imaging techniques, including brightfield, phase contrast, and fluorescence microscopy.
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2 protocols using ti e inverted light microscope
1
Wound Healing Assay in Primary Keratinocytes
2
Multimodal Imaging of Cellular Processes
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Confocal images were obtained with an LSM 510 (63× Plan-Apochromat objective; fixed imaging; ZEISS) or LSM 710 (63× Plan-Apochromat objective; ZEISS) using Argon2, HeNe, and HeNe2 lasers (live imaging) and Zen software (ZEISS). TIRF images were obtained with a TiE inverted light microscope (Nikon) equipped with a perfect focus, TIRF illuminator (Nikon), a 100×/1.49 NA TIRF objective (used in combination with a 1.5× optivar), Neo 5.5 cMOS camera (Andro), Sapphire 488- and 561-nm 50-mW lasers (Coherent, Inc.), and Elements software (Nikon).
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