HL-60 cells were infected with FAM-SE labeled pneumococcal strains as described before [26 (link)]. DNA was stained with Hoechst (Invitrogen) whereas actin cytoskeleton was stained with rhodamine-phalloidin (Invitrogen) [26 (link)]. Phagocytosis was analyzed with a Leica spectral SP5 confocal microscope using the Leica software (LAS-AF). Biofilm formation was observed using a Leica TCS-SP2-AOBS-UV confocal microscope. Briefly, pneumococcal isolates were grown on glass-bottom dishes (WillCo-dish, WillCo Wells) for 6 h at 34°C and stained with the BacLight kit showing living (green fluorescence) and dead (red fluorescence) bacteria, as previously described [29 (link)].
Tcs sp2 aobs uv confocal microscope
Manufactured by Leica
The TCS-SP2-AOBS-UV confocal microscope is a high-performance research tool designed for advanced microscopy applications. It features a tunable laser system and an acousto-optical beam splitter (AOBS) for versatile fluorescence excitation and detection. The microscope is equipped with ultraviolet (UV) laser capabilities, enabling the visualization and analysis of a wide range of samples.
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Lab products found in correlation
Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions)
Sp5 spectral confocal microscope, by Leica (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Matrigel coated coverslips, by BD (1 mentions)
Ix50 inverted system microscope, by Olympus (1 mentions)
Fluorescence mounting medium, by Agilent Technologies (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
2 protocols using tcs sp2 aobs uv confocal microscope
1
Pneumococcal Phagocytosis and Biofilm Imaging
2
Immunofluorescence Imaging of Cells on Matrigel
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Cells directly attached onto Matrigel-coated coverslips (BD Biosciences), were fixed with 1% paraformaldehyde (Sigma-Aldrich) in PBS containing 0.1% Triton X-100. Cells were washed three times with PBS and incubated with the corresponding antibodies and 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich). After incubation, samples were mounted with Mounting Fluorescence Medium (Dako), and images were captured using a TCS-SP2-AOBS-UV confocal microscope (Leica) with a ×63 oil-immersion objective. Displayed images were captured at the same sections in the different samples. Bright-field images were obtained using an IX50 inverted system microscope (Olympus), with LCPLFL ×20 and ×40 objectives. Cell and nucleus diameter and area were measured using ImageJ software.
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