An image from the slice by slice reconstruction of a double tilt series of tomograms of a Caulobacter Crescentus specimen. This image has been downsampled from the raw data image which can be accessed using the link provided to the Cell Centered Database.
Caulobacter cultures (strain 3724) were cryofixed using a cocktail of 2% Osmium tetroxide, 0.5% uranyl acetate in anhydrous acetone. High Pressure Freezing (HPF) using Balzers HPM 010 in 100 micron deep gold plated copper planchettes. Bacteria was centrifuged decanted and paced applied to planchettes. Freeze-substitution was conducted using this time and temperature schedule: -90 Celsius for 72 hours, raised 2 Celsius per hour for 15 hours, held at -60 Celsius for 12 hours, raised 5 Celsius per hour for 12 hours, held at -20 Celsius. Film images were acquired using a JEOL 4000, at a magnification of 30,000X, 400.0 Kv.