Coumarin 6 was utilized to visualize encapsulation into the shell of PBCA MB. A Leica TCS SP8 X inverted confocal microscope (Leica Microsystems, Wetzlar, Germany) equipped with a plan-apochromat 100x/1.40 oil-immersion objective was used to visualize coumarin 6-loading in PBCA MB. The samples were applied on a high precision cover glass (170 μm, No. 1.5H) from Marienfeld (Lauda-Königshofen, Germany). The excitation wavelength was set at λ = 470 nm via a filtered white light laser and the resulting emission was detected at λ = 491 - 556 nm. Deconvolution of the images was processed using the Huygens Professional software via the Classic Maximum Likelihood Estimation (CMLE) method.
Tcs sp8 x inverted
Manufactured by Leica
Sourced in Germany
The Leica TCS SP8 X inverted is a confocal laser scanning microscope designed for advanced imaging applications. It features a fully motorized and automated platform with a built-in laser system for high-resolution imaging of a wide range of samples.
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2 protocols using tcs sp8 x inverted
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Coumarin 6 Visualization of PBCA Microbubbles
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Cell Imaging and Staining Protocols
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Cells were stained with Calcein AM (cytoplasm) and Hoechst 33342 (nuclei) according to the manufacturer’s protocol. Briefly, the stock staining solution was made by mixing culture media DMEM without FBS (3 mL) and Hoechst 33342 (3 L), and adding Calcein AM solution (3 L for L-929, B16F10, CT26 cell lines; 0.5 L for THP-1, PBMC). Platelets were stained with Nile red as described above. Cell imaging was performed during the first hour (every 5 min), at 2 and 24 h.
CLSM was performed with Leica TCS SP8 X inverted confocal microscope (Leica Microsystems, Wetzlar, Germany) equipped with diode (405 nm) and argon laser sources. All measurements were performed using 20×/0.70 N.A. objective. The following settings were used: 405 nm excitation with 415–477 nm detection (blue channel, Hoechst 33342, cell nuclei), 488 nm excitation with 500–550 nm detection (green channel, Calcein AM, cell cytoplasm), and 514 nm excitation with 605–700 nm detection (red channel, Nile red, platelets).
CLSM was performed with Leica TCS SP8 X inverted confocal microscope (Leica Microsystems, Wetzlar, Germany) equipped with diode (405 nm) and argon laser sources. All measurements were performed using 20×/0.70 N.A. objective. The following settings were used: 405 nm excitation with 415–477 nm detection (blue channel, Hoechst 33342, cell nuclei), 488 nm excitation with 500–550 nm detection (green channel, Calcein AM, cell cytoplasm), and 514 nm excitation with 605–700 nm detection (red channel, Nile red, platelets).
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