Sporulating cells were concentrated by centrifugation at 8, 000 rpm for 1 min and immobilized on 2% agarose pads. Fluorescence microscopy was performed using an Olympus BX61 microscope equipped with a UplanF1 100X phase contrast objective lens and a CoolSnapHQ digital camera (Photometrics) or a Nikon TE2000 inverted microscope with a Nikon CFI Plan Apo VC 100X objective lens. Images were acquired using Metamorph software (Molecular DEVICES, Sunnyvale, CA, USA). Membranes were stained with TMA-DPH (50μM) (Molecular Probes; ThermoFisher Scientific) and fission of mother cell membranes was assessed as previously described (Doan et al., 2013 (link)). Image analysis and processing was performed in Metamorph.
Cfi plan apo vc 100x
Manufactured by Nikon
The CFI Plan Apo VC 100X is a high-quality microscope objective lens manufactured by Nikon. It is designed to provide excellent optical performance and clarity. The lens features a numerical aperture of 1.40 and a working distance of 0.13 mm.
Automatically generated - may contain errors
Lab products found in correlation
Metamorph software, by Molecular Devices (1 mentions)
Bx61 microscope, by Olympus (1 mentions)
Te2000, by Nikon (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Piezo z motor, by Nikon (1 mentions)
Orcaflash 4.0 camera, by Nikon (1 mentions)
Nile red, by Thermo Fisher Scientific (1 mentions)
Eclipse ti2, by Nikon (1 mentions)
2 protocols using cfi plan apo vc 100x
1
Sporulation Cell Membrane Fission Analysis
2
Multi-color Fluorescence Microscopy of Bacterial Cells
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Cells were collected by centrifugation at 3,300 ×g for 2 min, then immobilized on PBS or M9 agarose pads (2% wt/vol). Cell walls were stained with FDAAs (HADA or sBADA) (Tocris) at 100 μM. Cell membranes were stained with 1 μg/ml Nile Red (ThermoFisher). DNA was stained with DAPI (Molecular Probes) at 2 μg/ml, or with Propidium Iodide (Molecular Probes) at 5 μM. Standard-resolution fluorescence microscopy was performed using a Nikon Eclipse Ti2 or Ti inverted microscope with a Nikon CFI Plan Apo VC 100X objective lens. 3D-SIM images were acquired with a Nikon Ti2 inverted microscope equipped with an N-SIM Spatial Light Modulator, a Physik Instrument Piezo Z motor, Nikon Laser Illuminators, and captured with a Dual Hamamatsu Orca Flash 4.0 camera using Nikon Elements 5.11 acquisition software.
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