Twelve-day old neurons grown on 25 mm coverslips (Fisherbrand, Pittsburg, PA) were transduced with AdNull, Ad-Tat or Ad-U1. Forty-eight hours after treatment, cells were incubated with mitochondrial oxygen free radical indicator MitoSOX red (Life Technologies, Eugene, OR) for 30 min at 37 °C. Coverslips were mounted for confocal imaging in an open perfusion microincubator (PDMI-2; Harvard Apparatus). Images were obtained at 561 nm excitation by using a 63× oil objective of Confocal microscope (810; Carl Zeiss, Inc.).
25 mm coverslips
Manufactured by Thermo Fisher Scientific
25 mm coverslips are thin, circular glass or plastic pieces used to cover and protect samples on microscope slides. They provide a clear, transparent surface for observing and analyzing samples under a microscope.
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2 protocols using 25 mm coverslips
1
Mitochondrial ROS Detection in Transduced Neurons
2
Imaging Cellular Structures with Confocal Microscopy
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Brightfield images and overlay GFP images were captured on the Biotek Cytation 5. For confocal microscopy imaging, cells were seeded in 6-well culture plates (Corning) containing 25 mm coverslips (Fisher Scientific). Nuclei were stained with 50 μL/coverslip anti-fade DAPI (Invitrogen) and stained with 0.1 µg mouse α-tubulin IgG primary antibody (Santa Cruz Biotechnology and 0.5 μg Alexa Fluor 568-conjugated goat anti-mouse IgG secondary antibody (Invitrogen) per coverslip. Images were acquired with a Zeiss LSM 710 confocal microscope at 100X magnification (Carl Zeiss AG, Oberkochen, GER).
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