Cfi plan apo vc 100x objective

Sporulating cells were concentrated by centrifugation at 8 krpm 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 and a CoolSnapHQ digital camera (Photometrics) or a Nikon TE2000 inverted microscope with a Nikon CFI Plan Apo VC 100X objective. Images were acquired using Metamorph software. Membranes were stained with TMA-DPH (50 μM) (Molecular Probes) and fission of mother cell membranes was assessed as previously described [34 (link)]. Exposure times were 400 ms for TMA-DPH and mCherry, 200 ms for YFP and CFP, and 1,000 ms for σ^E-gfp. Image analysis and processing were performed in Metamorph.

Cells were imaged on 2% agarose pads containing 1X M9 salts (unless indicated otherwise) using a Nikon TE2000 inverted microscope outfitted with a Nikon Intensilight illuminator, a Coolsnap HQ2 CCD camera from Photometrics, a Nikon CFI Plan Apo VC 100x objective (1.4 NA) for differential interference contrast (DIC) imaging or a CFI Plan Apo DM 100x objective (1.4 NA) for phase contrast imaging. Please see figure legends for growth conditions used for specific experiments. Filter cubes for fluorescence image acquisition were from Chroma. mCherry and GFP fluorescence images were taken using the ET-mCherry filter set (Chroma 49008) and ET-GFP filter set (Chroma 49002), respectively. Images were captured using Nikon Elements software, exported and cropped for figure preparation in MetaMorph (Molecular Devices).