Paraffin-embedded lung tissue blocks were baked on the hotplate at 75 °C for 20 min and deparaffinized in xylene. The slides were rehydrated from 100%, 90%, to 70% alcohol, and then to PBS. We performed the antigen unmasking using the retriever (Cat. # 62700–10, Electron Microscopy Sciences) with R-Buffer A pH 6.0 (Cat. # 62706–10, Electron Microscopy Sciences) for 2 h to complete the cycle and cool down. Slides were blocked with 20% normal goat serum (NGS) in PBST for 2 h at room temperature. Slides were incubated with an anti-IL-8 antibody (Cat. # 550419, BD Pharmingen™) in 5% NGS with PBS at 4°C for overnight. Slides were washed with PBST and incubated with Alexa 488 coated goat anti-mouse antibody in 5% NGS/PBS for 2 h at room temperature. Slides were washed with PBST and stained with Hoechst (1:5000 in PBS, Invitrogen). Coverslips were mounted on slides using ProLong Glass Antifade Mountant (Cat. # P36982, Invitrogen) and dried out in the dark overnight. Confocal images were acquired using the ZEISS LSM 700 Upright laser scanning confocal microscope and ZEN imaging software (ZEISS).
Lsm 700 upright laser scanning confocal microscope
Manufactured by Zeiss
The ZEISS LSM 700 Upright laser scanning confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It utilizes laser illumination and a scanning mechanism to capture detailed, high-resolution images of samples. The system is suitable for a wide range of specimen types and can be configured with various objective lenses and detection options to meet the specific needs of researchers and scientists.
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2 protocols using lsm 700 upright laser scanning confocal microscope
1
Immunofluorescent Staining of IL-8 in Lung Tissue
2
Immunofluorescence Staining of IL-8 in Lung Tissue
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Paraffin-embedded lung tissue blocks were baked on the hotplate at 75°C for 20 min and deparaffinized in xylene. The slides were rehydrated from 100%, 90%, to 70% ethanol and then to PBS. We performed the antigen unmasking using the retriever (Cat. # 62700-10, Electron Microscopy Sciences) with R-Buffer A pH 6.0 (Cat. # 62706-10, Electron Microscopy Sciences) for 2 h to complete the cycle and cool down. Slides were blocked with 20% normal goat serum (NGS) in PBST for 2 h at room temperature. Slides were incubated with an anti-IL-8 antibody (Cat. # 550419, BD Pharmingen™) in 5% NGS with PBS at 4°C overnight. Slides were washed with PBST and incubated with Alexa 488 coated goat anti-mouse antibody in 5% NGS/PBS for 2 h at room temperature. Slides were washed with PBST and stained with Hoechst (1:5000 in PBS, Invitrogen). Coverslips were mounted on slides using ProLong Glass Antifade Mountant (Cat. # P36982, Invitrogen) and dried out in the dark overnight. Confocal images were acquired using the ZEISS LSM 700 Upright laser scanning confocal microscope and ZEN imaging software (ZEISS).
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