Recovered cells were fixed in 3% (v/v) glutaraldehyde in PBS for 2 hours at RT. Cells were pipetted and rinsed in the fixative during the fixation period. They were then washed 3 times in PBS, 10 min each, dehydrated in an ethanol series, treated for the methylation-acetylation procedure70 (link) and finally embedded in LR White resin (London Resin, EMA, UK). From resin-embedded blocks, semi-thin (2 µm thick) sections were visualized by phase contrast microscope (Leica DM2500) and imaged with a Leica DFC320 CCD. The images obtained were processed for quantitative studies using the software programs QWin Standard image analysis (Leica Microsystems) and Image J2.0 (http://www.imagejdev.org ). A sample of 10 cells per image were counted for the quantitative studies in three different replicates.
Qwin standard image analysis
Manufactured by Leica
QWin Standard is a comprehensive image analysis software solution developed by Leica. It provides users with a range of tools for the acquisition, processing, and analysis of microscopic images. The software's core function is to enable users to efficiently and accurately measure, quantify, and extract data from digital images obtained through various microscopy techniques.
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Lab products found in correlation
2 protocols using qwin standard image analysis
1
Quantitative Microscopy Analysis of Fixed Cells
2
Ultrastructural Analysis of Cell Samples
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Samples were fixed using 3% (v/v) glutaraldehyde in PBS for 2 hours at RT. Cells were pipetted and rinsed in the fixative during the fixation period. They were then washed three times with phosphate-buffered saline (PBS), 10 min each, dehydrated in an ethanol series, treated for the methylation-acetylation procedure (Testillano, González-Melendi, Ahmadian & Risueño, 1995) and finally embedded in LR White resin (London Resin, EMA, UK). From resin-embedded materials, semi-thin 2 µm thick sections were obtained from the LR White blocks, visualized by phase contrast in a Leica DFC320 microscope equipped with a Leica DM2500 CCD digital camera. Ultrathin sections were stained for 30 min with 5% (w/v) uranyl acetate and for 90 seconds with 0.3% (w/v) lead citrate, separated by a wash in distilled water. Samples were observed using a JEOL 1230 transmission electron microscope operating at 100 kV acceleration voltage. The images obtained were processed for quantitative studies using QWin Standard image analysis (Leica Microsystems) and Image J 2.0 (imagejdev.org) software.
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