Confocal microscopy was performed using the Leica TCS SP8 MP confocal microscope system (Germany). The primary antibodies included PTGDS (sc-390717, Santa Cruz Biotechnology), MYH9 (60233-1-Ig; 11128-1-AP, Proteintech Group), GSK3-β (22104-1-AP, Proteintech Group), p-H2AX (Ser139, 9718, CST), His (12698, CST) and Flag (14793, CST).
Tcs sp8 mp confocal microscope system
Manufactured by Leica
Sourced in Germany
The Leica TCS SP8 MP confocal microscope system is a high-performance imaging solution designed for advanced microscopy applications. It features a state-of-the-art laser scanning confocal architecture, enabling high-resolution, multi-dimensional imaging of samples. The system is capable of providing accurate and reliable data acquisition, making it a valuable tool for researchers and scientists across various fields of study.
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4 protocols using tcs sp8 mp confocal microscope system
1
Confocal Microscopy Analysis of Protein Targets
2
Quantification of DNA Damage in DLBCL
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DLBCL cells with indicated treatment were transferred to a glass slide using cytospin. 4% formaldehyde fixation was applied for 15 min and DLBCL cells were permeabilized using 0.1% Triton X 100 for 10 min After blocked with 5% goat serum for 1 h, slides were incubated with the primary antibody (p-H2AX, Ser139, 9718, CST) at 4 °C overnight. Slides were further incubated with secondary antibodies and DAPI. Leica TCS SP8 MP confocal microscope system (Germany) was used for confocal microscopy.
3
Immunofluorescence Analysis of DLBCL Cells
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DLBCL cells were transferred to slides by cytospin centrifugation, fixed in 4% paraformaldehyde, and permeabilized in 0.5% Triton X-100. Subsequently, slides were blocked with 5% normal goat serum for 1 h and incubated with primary antibodies at 4 °C overnight. The next day, the cells were washed and incubated with the secondary antibody (Invitrogen) for 1 h before mounting with DAPI. The Leica TCS SP8 MP confocal microscope system (Germany) was used to analyze images. The primary antibodies included CHST11 (Invitrogen, PA5-103698), MOB1B (ORIGENE, TA501388S), and YAP (Proteintech,13584-1-AP).
4
Immunofluorescence Imaging of Subcellular Localization
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Cells were washed three times with phosphate-buffered saline (PBS) and fixed with 4% w/v paraformaldehyde for 15 min at 4°C, followed by permeabilization with absolute methanol for 2 min at room temperature. Primary antibodies against indicated proteins and corresponding secondary antibodies were used to label the cells. To visualize the nuclei, cells were stained with 4,6-diamidino-2-phenylindole (DAPI; Sigma) or Hoechst 33342 (Thermo Fisher Scientific). Images were viewed either with a Zeiss Axiovert 200 M fluorescence microscope (with 63× objective) or Leica TCS SP8 MP confocal microscope system (with 63× objective). Quantification of fluorescence signals in different subcellular compartments was conducted using our previously described method (Xu et al., 2008 (link); Tong et al., 2006 (link)).
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