Adipogenesis was assessed by staining with Biodipy 493/503 (Invitrogen catalog no. D3922) for lipid droplet accumulation and by staining with Hoechst 33342 (Thermo Fisher catalog no. 62249) for nucleus number at 4 to 6 days post-induction (mouse cells) and 14 to 18 days post-induction (human cells) in individual wells of a 384-well plate (Sigma-Aldrich catalog no. CLS3770). The cells were imaged on a Keyence inverted fluorescence microscope (BZX-710) by using DAPI (excitation, 360/40 nm; emission, 460/50 nm; Keyence, OP-87762) and green fluorescent protein (excitation, 470/40 nm; emission, 525/50 nm; Keyence, OP-87763) filters. Individual wells were imaged in their entirety at 20× magnification to capture a 7-by-7 tiled/stitched grid (mouse studies) or at 10× magnification to capture a 5-by-5 tiled/stitched grid (human studies).
Inverted fluorescence microscope bzx 710
Manufactured by Keyence
The Inverted fluorescence microscope (BZX-710) is a laboratory equipment designed for imaging and analysis of fluorescent samples. It features an inverted design, allowing for easy access and manipulation of the specimen. The microscope is equipped with various fluorescence illumination and detection capabilities to enable visualization and study of fluorescently labeled biological specimens.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Biodipy 493 503, by Thermo Fisher Scientific (1 mentions)
Op 87763, by Keyence (1 mentions)
Op 87762, by Keyence (1 mentions)
Mf20 antibody, by Developmental Studies Hybridoma Bank (1 mentions)
2 protocols using inverted fluorescence microscope bzx 710
1
Adipogenesis Quantification via Fluorescence
2
Immunostaining of Human Muscle Myotubes
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Immunostaining of human muscle myotubes was performed as described previously.14 (link) Myoblasts were differentiated into myotubes for 6 days, and then were fixed with 4% paraformaldehyde (PFA). Myotubes were treated with 0.1% Triton X-100 in PBS for 10 min. After blocking with 2% BSA in PBS for 30 min, the expression of pan myosin heavy chain (MHC) in myotubes was detected with the MF-20 antibody (Developmental Studies Hybridoma Bank) followed by secondary antibody (Alexa Fluor 561 goat anti–mouse IgG1 1:1000; Invitrogen). Nuclei were counterstained with DAPI (Thermo Fisher Scientific). Representative images of cells were taken on a Keyence inverted fluorescence microscope (BZX-710).
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