Confocal laser scanning microscopy (CLSM) was used to observe the morphology and deposition mechanism of TE-NEs and WCS-TE-NEs. Briefly, 3 µL of Nile Red O fluorescent dye solution (0.02%, w/v) was added to 500 µL of the TE-NEs to stain the MCT oil. As an amine-reactive fluorescent dye, fluorescein isothiocyanate (FITC) was used to stain WSC. The synthesis of FITC-labeled WSC was based on the reaction between isothiocyanate group of FITC and the amino group of chitosan [17 (link)]. WSC was stained by mixing WSC with 0.05% (w/v) FITC solution in the dark at ambient temperature for 4 h. The FITC-labeled WSC was precipitated by centrifuge at 10,000 rpm for 30 min and the unreacted FITC was separated until no fluorescence was detected in the supernatant. CLSM 700 confocal microscope (Carl Zeiss, Oberkochen, Germany) was used to analyze the samples at a magnification of 40×. Nile Red O and FITC dyes were excited at 543 and 488 nm, respectively. The z-stack images were obtained using the LSM 700 ZEN software.
Clsm 700 confocal microscope
Manufactured by Zeiss
Sourced in Germany
The CLSM 700 is a confocal laser scanning microscope manufactured by Zeiss. It is designed to produce high-resolution, three-dimensional images of samples by scanning the specimen with a focused laser beam and detecting the emitted fluorescence or reflected light. The CLSM 700 features superior optical performance and advanced imaging capabilities for a wide range of applications in life sciences and materials research.
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Lab products found in correlation
Rabbit anti mouse cd8α d4w2z, by Cell Signaling Technology (2 mentions)
Anti mouse cxcl5, by LSBio (2 mentions)
Nexgen decloaker chamber, by Biocare Medical (2 mentions)
Warp red chromagen kit, by Biocare Medical (2 mentions)
Rabbit anti mouse β catenin, by Cell Signaling Technology (2 mentions)
Rat anti mouse ly6g, by Abcam (2 mentions)
Vina green chromogen kit, by Biocare Medical (2 mentions)
Pd l1, by Abcam (1 mentions)
4 protocols using clsm 700 confocal microscope
1
Visualizing Nanoparticle Deposition Mechanisms
2
Comprehensive Immunohistochemistry Analysis
3
Comprehensive Immunohistochemistry Analysis
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Tissues were paraffin embedded and processed following standard protocols and imaged with a Zeiss CLSM 700 confocal micro-scope. For Lung and Tumor H&E, hematoxylin (VWR, 95057–844) was stained followed by Eosin (VWR, 95057–848). The following antibodies were used for Immunohistochemistry: Rabbit anti-mouse CD8α (D4W2Z) 1:400 (Cell Signaling, 98941), anti-mouse CXCL5 1:200 (LsBio, LS-C104413), rat anti-mouse Ly6g 1:100 (Abcam, ab25377), rabbit anti-mouse β-catenin 1:100 (Cell signaling, 8480). Antigen retrieval was performed by incubation in BioCare’s Nexgen Decloaker chamber for 20 minutes at 90°C. Anti-rabbit polymers were used as secondary antibodies and Vina Green Chromogen Kit (Biocare Medical, BRR 807 AH) was used as substrate for CD8. For CXCL5, LY6G, and β-catenin, anti-rat or rodent polymers were used as secondary antibodies, and Warp Red Chromagen Kit (Biocare Medical, 901-WR806–081017) was used as substrate. ImageJ software was used for quantification.
4
Immunohistochemical Staining of Paraffin-Embedded Tissues
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