Brains of 7 day old female and male flies were dissected in PBS and directly transferred to 4% PFA with 0.1% Triton-X (PBST) for fixation over 2 hours on ice. After three 15 minute washes with PBST and blocking in 5% normal goat serum (NGS) in PBST, the primary antibodies mouse anti-brp (1:30, Developmental Studies Hybridoma Bank) and rabbit anti-GFP (1:500, Thermo Fisher) were applied in PBST with 5% NGS for 4 days nutating at 4°C. Next, the brains were washed four times for 15 minutes with PBST, blocked again with 5% NGS in PBST and incubated with the secondary antibodies goat anti-mouse Alexa Fluor 633 and goat anti-rabbit Alexa Fluor 488 (1:250 each, Thermo Fisher) in PBST with 5% NGS for 5 days nutating at 4°C. After four final washing steps with PBST, brains were mounted in VectaShield (Vector Labs). Stacks of immunostained brains were scanned on a Zeiss cLSM 880 confocal microscope with a z-step size of 0.44 μm. The confocal stacks were manually reconstructed as label fields in AMIRA (6.7, Thermo Fisher) and identified on the basis of the published Drosophila melanogaster atlases [51 (link)]. Surface renders were created and smoothed in AMIRA with the surface gen and smooth surface tools.
Clsm 880 confocal microscope
Manufactured by Zeiss
Sourced in Germany
The CLSM 880 is a confocal laser scanning microscope from Zeiss. It is designed to provide high-resolution optical imaging of samples by scanning the specimen with a focused laser beam and collecting the emitted fluorescence or reflected light. The core function of the CLSM 880 is to enable detailed observation and analysis of microscopic structures and processes.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 633 goat anti mouse, by Thermo Fisher Scientific (2 mentions)
Vectashield, by Vector Laboratories (2 mentions)
Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (2 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (2 mentions)
Mouse anti brp, by Developmental Studies Hybridoma Bank (2 mentions)
3 protocols using clsm 880 confocal microscope
1
Drosophila Brain Immunostaining and Confocal Imaging
2
Confocal Imaging of Aerosol Fiber Samples
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Confocal microcopy imaging was carried out on a Zeiss CLSM 880 confocal microscope (Germany). To visualize the VSPs and optionally trace the liquid water content on the fibers, the fiber sample was amount on a glass slide and sealed with a coverslip. To estimate the size of the aerosols, the solution was sprayed on a Petri dish with hydrophobic surface and imaged by a confocal microscope immediately. The AGA and NR dyes were excited with the laser wavelengths of 405 nm and 488 nm, respectively.
3
Drosophila Neuroanatomy Mapping
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Brains of 7 day old female and male flies were dissected in PBS and directly transferred to 4% PFA with 0.1% Triton-X (PBST) for fixation over 2 hours on ice. After three 15 minute washes with PBST and blocking in 5% normal goat serum (NGS) in PBST the primary antibodies mouse anti-brp (1:30, Developmental Studies Hybridoma Bank) and rabbit anti-GFP (1:500, Thermo Fisher) were applied in PBST with 5% NGS for 4 days nutating at 4°C. Next, the brains were washed four times for 15 minutes with PBST, blocked again with 5% NGS in PBST and incubated with the secondary antibodies goat anti-mouse Alexa Fluor 633 and goat anti-rabbit Alexa Fluor 488 (1:250 each, Thermo Fisher) in PBST with 5% NGS for 5 days nutating at 4°C. After four final washing steps with PBST, brains were mounted in VectaShield (Vector Labs). Stacks of immunostained brains were scanned on a Zeiss cLSM 880 confocal microscope with a z-step size of 0.44 µm. The confocal stacks were manually reconstructed as label fields in AMIRA (6.7, Thermo Fisher) and identified on the basis of the published Drosophila melanogaster atlases [51] .
Surface renders were created and smoothed in AMIRA with the surface gen and smooth surface tools.
Surface renders were created and smoothed in AMIRA with the surface gen and smooth surface tools.
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