For immunofluorescence staining, the cells were incubated with primary antibodies against VE-cadherin (Cell Signaling Technology [CST], USA), vimentin (CST) or fibronectin (Abcam), followed by incubation with Alexa Fluor 488- or 555-conjugated secondary antibodies (Invitrogen, USA). For confocal microscopy, the cells grown on coverslips were counterstained with DAPI and were imaged using a confocal laser-scanning microscope (Carl Zeiss, Germany) with a core data acquisition system (Applied Precision).
Core data acquisition system
Manufactured by Cytiva
Sourced in Germany
The Core data acquisition system is a compact and versatile instrument designed for real-time data collection and analysis. It features high-speed data acquisition capabilities, allowing for the capture and processing of various sensor inputs with precision and reliability.
Automatically generated - may contain errors
Lab products found in correlation
Confocal laser scanning microscope, by Zeiss (2 mentions)
Fibronectin, by Abcam (1 mentions)
Alexa fluor 488 or 555 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Ve cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Abcam (1 mentions)
Lsm 700 laser scanning microscope, by Zeiss (1 mentions)
Fluorescently conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti human cd8, by Abcam (1 mentions)
Anti p65, by Cell Signaling Technology (1 mentions)
Leica tcs sp8, by Leica camera (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions)
Alexa fluor 488 donkey anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Mab394, by R&D Systems (1 mentions)
Alexa fluor 555 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
A confocal laser scanning microscope, by Zeiss (1 mentions)
Gpr37, by Abcam (1 mentions)
Coverslip, by Corning (1 mentions)
Leica tcs sp8, by Leica Microsystems (1 mentions)
6 protocols using core data acquisition system
1
Immunofluorescence Imaging of Cell Markers
2
Multicolor Immunofluorescence Staining
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For immunofluorescence staining, the specimens were incubated with mouse anti-hCD68 mAb (diluted 1:100), rabbit anti-hIL-6 Ab (diluted 1:100) and rabbit anti-hIL-8Ab (diluted 1:100) at 4°C overnight. Secondary staining with Alexa-Fluor-555 conjugated donkey anti-rabbit and Alexa-Fluor-488 conjugated goat anti-mouse secondary antibodies was carried out at room temperature for 60 min, followed by DAPI nuclear counterstaining for 10 min. Images were taken with a Zeiss LSM 700 laser scanning microscope (Carl Zeiss) with a core data acquisition system (Applied Precision). For control experiments, primary antibody was substituted with normal rabbit serum.
3
Immunofluorescence Staining of CD8 and NF-κB
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Cells were fixed with 4% paraformaldehyde for 15 min at room temperature and permeabilized using 0.2% Triton X-100. Frozen sections were obtained from fresh tissues after surgery. Paraffin-embedded tumor sections were deparaffinized and antigen repaired. Afterwards, the tissue sections or cell-adherent slides were blocked with 10% bovine serum albumin (BSA; Sigma) for 1 h and incubated with anti-human CD8 (Abcam) and anti-p65 (Cell Signaling Technology). After rigorous washing with PBS, sections or slides were incubated with fluorescently conjugated secondary antibodies (Thermo Fisher Scientific). Isotype matched antibodies were used as controls. After counterstaining with DAPI (Abcam), we acquired images using a confocal laser-scanning microscope (Leica TCS-SP8) with a core data acquisition system (Applied Precision).
4
Immunofluorescence Analysis of CCL18 and PITPNM3
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Naive CD4+ T cells isolated from PB of healthy donors were incubated with primary antibodies against CCL18 (Cat# MAB394, mouse anti-human, R&D; 20 μg/ml) or PITPNM3 (Cat# NBP1-31070, rabbit anti-human, Novus; 1:100), followed by incubation with Alexa Fluor 488 donkey anti-mouse IgG (H+L) and Alexa Fluor 555 goat anti-rabbit IgG (H+L) (Thermo Fisher). For confocal microscopy, the cells on cover slips were counterstained with DAPI and imaged using a confocal laser-scanning microscope (Carl Zeiss) with a core data acquisition system (Applied Precision).
5
Immunofluorescent Staining and Confocal Microscopy
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For immunofluorescent staining, the cells were incubated with primary antibodies against REG4 (R&D) and GPR37 (Abcam), followed by incubation with iFluor 594 anti-rabbit IgG antibody (AAT Bioquest) or Alexa488-conjugated anti-goat IgG antibody (Life technologies). For confocal microscopy, the cells on cover-slips were counterstained with DAPI and imaged using a confocal laser-scanning microscope (Carl Zeiss) with a core data acquisition system (Applied Precision).
6
Immunocytochemistry of Cytoskeletal Proteins
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The 4% paraformaldehyde was used to fix the cells on the coverslips (Corning) for 15 min at room temperature (RT). Afterward, 10% bovine serum albumin (BSA, Sigma) was used to block these cells for 1 h. Specific primary antibodies against MAP7D2, MYH9, and HMGB1 were used to incubate cells at 4°C overnight. Isotype matched antibodies were used as negative control experiments. After washing with Tris-buffered saline with 0.1% (v/v) Tween 20 (TBST) 3 times, the slides were probed with Alexa Fluor 594 donkey anti-rabbit IgG antibodies and Alexa Fluor 488 donkey anti-mouse IgG for 1 h at RT. The samples were then mounted with DAPI and the confocal laser-scanning microscope (Leica TCS-SP8, Leica Microsystems) was used to capture the images with a core data acquisition system (Applied Precision).
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