A549, NCI-H1650, HCC827, and NCI-H358 cells were cultured on Falcon chamber slides (BD Biosciences, USA) until 50-60% confluence before being fixed with 4% paraformaldehyde and permeabilized with 0.3% Triton X-100. Cells were immersed three times with cold phosphate-buffered saline (PBS), incubated with FOXM1 (Santa Cruz Biotechnology, USA), E-cadherin, N-cadherin, vimentin (Abcam, Cambridge, UK), and SNAIL (Abgent, USA) primary antibodies at 4°C overnight, subsequently incubated with corresponding Alexa Fluor-conjugated secondary antibodies (Life Technologies, USA) at room temperature for 1 h, and mounted using ProLong® Gold Antifade Reagent with DAPI (Life Technologies, USA). Microscopic images of cells were obtained using a Leica inverted fluorescence microscope (Leica Microsystems, Wetzlar, Germany) with ProgRes Image Capture Software (JENOPTIK Optical Systems, Jena, Germany) and a Leica Confocal LAS-AF SP5 System (Leica Microsystems, Germany).
Confocal las af sp5 system
Manufactured by Leica
Sourced in Germany
The Leica Confocal LAS-AF SP5 System is a high-performance confocal laser scanning microscope. It is designed for advanced imaging, analysis, and documentation of biological samples. The system utilizes a multi-channel detection system and a range of laser lines to enable comprehensive imaging capabilities.
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Lab products found in correlation
Inverted fluorescence microscope, by Leica (2 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (2 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)
E cadherin, by Abcam (2 mentions)
N cadherin, by Abcam (2 mentions)
Progres image capture software, by Jenoptik (2 mentions)
Falcon chamber slides, by BD (1 mentions)
Snail, by Abcepta (1 mentions)
Foxm1, by Santa Cruz Biotechnology (1 mentions)
Vimentin, by Abcam (1 mentions)
2 protocols using confocal las af sp5 system
1
Immunofluorescence Analysis of EMT Markers
2
Immunofluorescence Assay of Epithelial and Neural Cadherins
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LoVo, HT29 and RKO cells were cultured until 50-60% confluence before being fixed with 4% paraformaldehyde and being permeabilized with 0.3% Triton X-100. Cells were washed three times in phosphate-buffered saline (PBS) and incubated with PAK7, epithelial cadherin (E-cadherin) and neural cadherin (N-cadherin) (Abcam, UK) primary antibodies at 4°C overnight then, incubated with corresponding Alexa Fluor-conjugated secondary antibodies (Life Technologies, USA) at room temperature for 1 h, and mounted using ProLong® Gold Antifade Reagent with DAPI (Life Technologies, USA). Microscopic images of the cells were obtained using a Leica inverted fluorescence microscope (Leica Microsystems, Wetzlar, Germany) with ProgRes Image Capture Software (JENOPTIK Optical Systems, Jena, Germany) and a Leica Confocal LAS-AF SP5 System (Leica Microsystems, Germany).
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