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Metamorph 7.5 imaging software

Manufactured by Molecular Devices
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Metamorph 7.5 is an imaging software developed by Molecular Devices. The software's core function is to capture, process, and analyze digital images from a variety of microscopy and imaging techniques.

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Lab products found in correlation

Orca flash4.0 lt digital cmos camera, by Hamamatsu Photonics (2 mentions) Oil objective, by Olympus (2 mentions) Lab tek 8 well chambers, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) X tremegene 9, by Roche (1 mentions) Leica sp5 inverted confocal microscope, by Leica camera (1 mentions) Axiovert 200 confocal microscope, by Zeiss (1 mentions) Epifluorescence microscope, by Olympus (1 mentions) Orca er ccd camera, by Hamamatsu Photonics (1 mentions) Zyla 4.2 scmos camera, by Oxford Instruments (1 mentions) Stage top incubation system, by Tokai Hit (1 mentions) Axiovert 200, by Zeiss (1 mentions) Nis elements software, by Nikon (1 mentions) Eclipse ti microscope, by Nikon (1 mentions) Ix71 microscope, by Olympus (1 mentions)

4 protocols using metamorph 7.5 imaging software

1

Ciliogenesis Induction in Mouse Cell Lines

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NIH-3T3, mIMCD3 and MEFs were cultured in DMEM (Gibco) supplemented with 10% FBS. For all transient transfections, cells were transfected with the respective DNA constructs by plating them directly in a transfection solution containing DNA plasmid and Xtremegene 9 (Roche). Cells were plated on poly(D-lysine)-coated borosilicate glass Lab-Tek 8-well chambers (Thermo Scientific). Ciliogenesis was induced by serum starvation for 24 h. Live cell imaging was mostly performed using an IX-71 (Olympus) microscope with a 40× oil objective (Olympus) (with additional 1.6× optical zoom) and a CoolSNAP HQ CCD camera (Photometrics). Micrographs were taken using Metamorph 7.5 imaging software (Molecular Devices).
Garcia-Gonzalo F.R., Phua S.C., Roberson E.C., Garcia G I.I.I., Abedin M., Schurmans S., Inoue T, & Reiter J.F. (2015). Phosphoinositides Regulate Ciliary Protein Trafficking to Modulate Hedgehog Signaling. Developmental cell, 34(4), 400-409.
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2

Live-cell Imaging of Organelles

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Live-cell imaging of ER, Golgi, and the plasma membrane was conducted using the Axiovert135TV epifluorescence microscope (Zeiss) with 63× oil objective (Zeiss) and Olympus epifluorescence microscope with 40× oil objective and an additional 1.6× magnification. Twenty-four hours after transfection, cells were imaged by the CCD camera (QImaging) driven by Metamorph 7.5 imaging software (Molecular Devices) at 15 second time interval for at least 15 minutes. Mitochondria were imaged on the Leica SP5 inverted confocal microscope, with resonant scanner, HCX PL APO cS 40× objective lens, NA = 1.25 at 15 second time interval for at least 15 minutes. Mitochondria were also imaged on the spinning-disc Axiovert 200 confocal microscope (Zeiss). YFP and mCherry excitations were conducted with an argon laser (CVI-Melles Griot) with 40× objective (Zeiss) and an additional 1.6× magnification and NA = 1.30. 100 nM of rapamycin (Tecoland) was added after approximately 2 min and 30 seconds of imaging.
Suarez A., Ueno T., Huebner R., McCaffery J.M, & Inoue T. (2014). Bin/Amphiphysin/Rvs (BAR) family members bend membranes in cells. Scientific Reports, 4, 4693.
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3

Quantitative Imaging of Membrane Contacts

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Most imaging was conducted on a IX71 microscope (Olympus) with a 63x objective (and 1.6x zoom) and an ORCA-Flash4.0 LT Digital CMOS camera (Hamamatsu). Rate constant of FKBP recruitment by sFRB1 was imaged at 10 s intervals with a spinning disc confocal, inverted Axiovert 200 (Zeiss) with a 40x objective and an Orca ER CCD camera (Hamamatsu). Both microscopes were driven by Metamorph 7.5 imaging software (Molecular Devices). Four-color imaging in Fig. 4 was conducted with an Eclipse Ti microscope (Nikon) with a 60x objective (and 1.5x zoom) and a Zyla 4.2 sCMOS camera (Andor), driven by NIS Elements software (Nikon). Unless otherwise indicated imaging was done at 1-3 min intervals for 12-30 mins, at times with between 3-5 0.5 um-spaced z positions. Images analyzed and shown are from a single plane, and not maximum intensity projections. Microscopy experiments applying CIT to membrane contact sites were conducted at 37°C, 5% CO2 and humidity with a stage top incubation system (Tokai Hit).
Wu H.D., Kikuchi M., Dagliyan O., Aragaki A.K., Nakamura H., Dokholyan N.V., Umehara T, & Inoue T. (2020). Rational design and implementation of a chemically inducible hetero-trimerization system. Nature methods, 17(9), 928-936.
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4

Live-cell Microscopy Imaging Protocol

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Live-cell imaging experiments were mostly performed using an IX-71 (Olympus) microscope with a 63x oil objective (Olympus) (with additional 1.6x optical zoom) and a Cool-SNAP HQ charge-coupled device camera (Photometrics) or an ORCA-Flash4.0 LT Digital CMOS camera (Hamamatsu). Time-lapse imaging experiments were performed with either 2-min or 5-min intervals, and between 3 and 11 0.5-µm z-stacks were taken at each time point. Images shown were mostly maximum intensity projection. Micrographs were taken and analyzed using MetaMorph 7.5 imaging software (Molecular Devices).
Phua S.C., Chiba S., Suzuki M., Su E., Roberson E.C., Pusapati G.V., Setou M., Rohatgi R., Reiter J.F., Ikegami K, & Inoue T. (2017). Dynamic remodeling of membrane composition drives cell cycle through primary cilia excision. Cell, 168(1-2), 264-279.e15.
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