Third-instar larval preparations (muscles 6/7) were filleted and fixed in 4% paraformaldehyde, washed, and incubated overnight at 4°C with primary antibodies. Secondary antibodies were applied at room temperature for 2 hr. The following primary antibodies were used: anti-Myc (9E10 Santa Cruz), anti-GFP 3E6 (1:500; mouse; Life Technologies), anti-NC82 (1:100; mouse; Developmental Studies Hybridoma Bank), anti-HA antibody (1:1000; Rabbit; cell signaling technology) and anti-DLG (1:10,000; rabbit). Alexa-conjugated secondary (488, 555) antibodies and Cy5-conjugated goat ant-HRP were used at 1:500 (Life Technologies; Molecular Probes). Larval preparations were mounted in Vectashield (Vector) and imaged with an Axiovert 200 (Zeiss) inverted microscope, a 100X Plan Apochromat objective (1.4 NA) and a cooled charge-coupled device camera (Coolsnap HQ, Roper). Slidebook 5.0 Intelligent Imaging Innovations (3I) software was used to capture, process and analyze images. Structured illumination microscopy imaging was performed using the N-SIM Nikon system, consisting of a Nikon Ti-E Microscope equipped with a Apo TIRF 100×/1.49 Oil objective and an Andor DU897 Camera.
Du897 camera
Manufactured by Nikon
The DU897 Camera is a professional-grade laboratory equipment designed for high-precision imaging applications. It features a high-resolution sensor, advanced optics, and robust construction to capture detailed and accurate images in controlled environments.
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Lab products found in correlation
Anti nc82, by Developmental Studies Hybridoma Bank (2 mentions)
Anti myc, by Santa Cruz Biotechnology (2 mentions)
Anti gfp 3e6, by Thermo Fisher Scientific (2 mentions)
Cy5 conjugated goat ant hrp, by Thermo Fisher Scientific (2 mentions)
Apo tirf 100 1.49 oil objective, by Nikon (2 mentions)
Axiovert 200, by Zeiss (2 mentions)
Vectashield, by Vector Laboratories (2 mentions)
Ti e microscope, by Nikon (2 mentions)
Anti dlg, by Developmental Studies Hybridoma Bank (2 mentions)
Goat anti rabbit alexa 488, by Thermo Fisher Scientific (1 mentions)
Goat anti mouse alexa 568, by Thermo Fisher Scientific (1 mentions)
Facl4, by Santa Cruz Biotechnology (1 mentions)
3 protocols using du897 camera
1
Larval Neuromuscular Junction Immunostaining
2
Larval Neuromuscular Junction Immunostaining
3
Immunofluorescence Imaging of ER Proteins
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Cells were grown on 14 mm round coverslips and fixed in 3% formaldehyde in PBS pH 7.4 containing 0.025% glutaraldehyde. Cells were permeabilized with 0.1% Triton and incubated with primary antibodies. Antibodies used were as follows: FACL4 (mouse monoclonal, 1:500, Santa Cruz Biotechnology Inc.); InsP3R type 3 (mouse monoclonal, 1:500, Becton Dickinson Biosciences); rodent Nox4 (rabbit polyclonal, 1:500; Anilkumar et al, 2008 ). Secondary antibodies were as follows: goat anti‐mouse Alexa 568 and goat anti‐rabbit Alexa 488 (both 1:500, Thermo Fisher). Cells were mounted in Mowiol and imaging was performed on an inverted Nikon Ti‐E microscope equipped with a Yokogawa CSU‐X1 spinning‐disk confocal unit, an Andor DU‐897 camera, a Sutter filter wheel, and Nikon 100 × 1.49 NA objective (Nikon Instruments).
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