Images of squashed preparations were collected with a x100 objective on a Leica DM6B epifluorescence microscope, this microscope was equipped with an ORCA-Flash4.0 V2 Digital CMOS camera (Hamamatsu) and controlled by the Metamorph software 7.7 (Molecular devices). Images of whole mount brain lobes were acquired on a Nikon A1R inverted TiE confocal microscope with a 40X 1.3NA or a X60 1.4 NA objective in NIS Element software. For the characterization of Spd2 and Fzr localization in interphase centrioles, images were acquired with a N-SIM Nikon microscope in 3D SIM mode before image reconstruction using the NIS Elements software (Gustafsson et al., 2008 (link)). The system is equipped with an APO TIRF SR 100x 1.49NA oil immersion, a laser illumination (488nm 200mW, 561nm 100mW, 640nm 100mW) and an EMCCD DU-897 Andor camera. Images were acquired with the following protocol, a Z stack (0.12 micron steps) was acquired. Images were then reconstructed using Nikon elements software.
Metamorph software 7
Manufactured by Molecular Devices
Sourced in United States, Japan
MetaMorph software 7.7 is a comprehensive software package designed for imaging and analysis of biological samples. It provides a suite of tools for acquiring, processing, and analyzing digital images from a variety of microscopy techniques. The software's core function is to enable users to capture, manage, and quantify data from their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Zyla 4.2 scmos camera, by Oxford Instruments (2 mentions)
Ti e inverted microscope, by Yokogawa (2 mentions)
Csu 22 spinning disk head, by Yokogawa (2 mentions)
Na plan apo oil immersion objective, by Nikon (2 mentions)
Csu w1 spinning disk system, by Nikon (2 mentions)
Na plan apo λ oil objective, by Nikon (2 mentions)
Nis elements 5.11.03 ar software, by Nikon (2 mentions)
γh2ax, by Merck Group (2 mentions)
Eclipse te2000, by Nikon (2 mentions)
Orca flash4.0 v2 digital cmos camera, by Hamamatsu Photonics (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Schneider s drosophila medium, by Thermo Fisher Scientific (1 mentions)
Ti e inverted microscope, by Nikon (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
P35g 1.5 14 c, by MatTek (1 mentions)
C9100 02 camera, by Hamamatsu Photonics (1 mentions)
Depex, by Avantor (1 mentions)
C9100 02 emccd camera, by Hamamatsu Photonics (1 mentions)
6 protocols using metamorph software 7
1
Multimodal Microscopy for Centriole Analysis
2
Imaging Drosophila Larval Brain Dynamics
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Mid third-instar larval brains were dissected in Schneider's Drosophila Medium (21720-024, Gibco) supplemented with 10% heat-inactivated fetal bovine serum (10500, Gibco), Penicillin (100 units ml−1) and Streptomycin (100 μg ml−1) (Penicillin-Streptomycin 15140, Gibco). Two to four brains were placed on a glass bottom 35 mm dish (P35G-1.5-14-C, MatTek Corporation) with 10 μl of medium, covered with a permeable membrane (Standard membrane kit, YSI) and sealed around the membrane borders with oil 10S Voltalef (VWR BDH Prolabo). Images were recorded using a Yokagawa CSU-X1 spinning head mounted on a Nikon TiE inverted microscope. The microscope was equipped with an EMCCD Evolve 512 × 512 (Photometrics) and controlled by the Metamorph software 7.7 (Molecular devices). Four-dimensional z-stacks of 10-30 μm at 1 μm intervals were acquired every 30 or 60 s using a 60 × NA 1.4 oil immersion objective. Images were processed with ImageJ.
3
Immunofluorescence Imaging of DNA Damage
4
Immunofluorescence Imaging of DNA Damage
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Immunofluorescence was performed as described28 (link),51 (link). Primary antibodies: γH2AX (1:400–500, MilliporeSigma, 05–636-I, clone JBW301), LBR (1:100, Abcam, ab32535, clone E398L). Secondary antibodies: Alexa Fluor, 488 (A11029), 568 (A11031) and 647 (A21236) (1:1000, Life Technologies). EdU was added 5 h before fixation.
Confocal images were collected using a Nikon Ti-E inverted microscope with a Yokogawa CSU-22 spinning disk head with the Borealis modification. Z-stacks were collected for 9 images at 0.4–0.6 μm spacing using a CoolSnap HQ2 CCD camera (Photometrics) and a 60x/1.40 NA Plan Apo oil immersion objective (Nikon) using Metamorph Software 7.10.2.240 (Molecular Devices). Alternatively, a Ti2 inverted microscope fitted with a CSU-W1 spinning disk system (Nikon) was used. Z-stacks were collected to cover the whole volume of cells at 0.4–0.6 μm spacing using a Zyla 4.2 sCMOS camera (Andor) and a 60x/1.40 NA Plan Apo λ oil objective and NIS-Elements 5.11.03 AR software (Nikon).
Confocal images were collected using a Nikon Ti-E inverted microscope with a Yokogawa CSU-22 spinning disk head with the Borealis modification. Z-stacks were collected for 9 images at 0.4–0.6 μm spacing using a CoolSnap HQ2 CCD camera (Photometrics) and a 60x/1.40 NA Plan Apo oil immersion objective (Nikon) using Metamorph Software 7.10.2.240 (Molecular Devices). Alternatively, a Ti2 inverted microscope fitted with a CSU-W1 spinning disk system (Nikon) was used. Z-stacks were collected to cover the whole volume of cells at 0.4–0.6 μm spacing using a Zyla 4.2 sCMOS camera (Andor) and a 60x/1.40 NA Plan Apo λ oil objective and NIS-Elements 5.11.03 AR software (Nikon).
5
Fluorescence Microscopy Imaging Protocol
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The tissue sections were imaged with a Nikon Eclipse TE2000 inverted epifluorescence microscope using Nikon objectives of 10x and 20x linear magnification with infinity correction. An X-Cite™ (Exfo, Vanier, QC, Canada) 120 W metal halide light source and a liquid light guide were used to illuminate the tissue samples. The excitation and emission filters (Chroma Technology, Bellows Falls, VT, USA) were controlled by a MAC5000 controller (Ludl, Hawthorne, NY, USA) and MetaMorph® software 7.8 (Molecular Devices, Downington, PA, USA). The fluorescence microscopy 16-bit fluorescent images were digitally recorded on a C9100-02 camera (Hamamatsu, Japan), digitally recombined and pseudocolored based on recording wavelength.
6
Tissue Clearing and Imaging Protocol
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