Computational protocol: Application of an E. coli signal sequence as a versatile inclusion body tag

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Protocol publication

[…] Unless stated otherwise, the following procedure was used to perform phase-contrast microscopy: Cells were first fixed by incubation in PBS containing 2.8% formaldehyde and 0.04% glutaraldehyde at room temperature for 15 min. The cells were kept cold during all subsequent procedures. To remove residual formaldehyde and glutaraldehyde, the cells were collected by low-speed centrifugation, transferred to a new tube and resuspended in fresh PBS. After three additional washings in PBS, the cells were resuspended in PBS before immobilization on 1% agarose in water slab-coated object glasses as described []. Samples were photographed with a CoolSnap fx (Photometrics) CCD camera mounted on an Olympus BX-60 fluorescence microscope through a UPLANFl 100 × /1.3 oil objective. Images were acquired with Micro-Manager ( with direct output of the desired hyperstack structure for ImageJ by Wayne Rasband ( […]

Pipeline specifications

Software tools μManager, ImageJ
Applications Phase-contrast microscopy, Microscopic phenotype analysis
Organisms Escherichia coli, Homo sapiens, Escherichia coli K-12
Diseases Drug-Related Side Effects and Adverse Reactions