The size of CFP-expressing S. flexneri infection foci formed in HT-29 cell monolayers was determined in 384-well plate format as described previously [20] (link). After fixation and DAPI staining, the plates were imaged using a TE2000 automated microscope (Nikon) equipped with a motorized stage (Prior), motorized filter wheels (Sutter Instrument, Inc.) and a 10× objective (Nikon) mounted on a piezo focus drive system (Physik Instrumente). Image acquisition was conducted using the Metamorph 7.1 Software (Molecular Devices, Inc.). For image analysis, high intensity pixels corresponding to the bacteria were selected against the low intensity pixels corresponding to the background levels by using the threshold function of the MetaMorph 7.1 imaging software. Selected pixels were clustered by proximity using the “close” morphological filter in order to create a single object that represented a given focus of infection. The integrated morphometry analysis (IMA) module was used to determine the area of each infection focus in a given image. Image analyses were conducted on at least 100 infection foci in three independent experiments.
Motorized filter wheels
Manufactured by Sutter Instruments
Motorized filter wheels are laboratory equipment designed to facilitate the automatic and precise control of optical filters in various scientific applications. The core function of motorized filter wheels is to provide a reliable and efficient method for rapidly and accurately positioning multiple optical filters into the light path, enabling seamless filter changes and efficient data acquisition.
Automatically generated - may contain errors
2 protocols using motorized filter wheels
1
Quantifying Shigella Infection Foci in HT-29 Cells
2
Automated High-Throughput Microscopy Imaging
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