Volume 9, Issue 1, January 2019

Visualization of Extracellular Trap by Light Microscopy

Author(s): Haritha V H, Binchu V Shaji and *Anie Y
Abstract: Aim:Neutrophil extracellular traps (N(ET)) are structures made of DNA studded with antimicrobial peptides, that are ejected out of many types of white blood cells including neutrophils. They are released as a defense strategy to trap/kill pathogens. NET-DNA is usually observed after staining with fluorescent stains specific to DNA or NET-proteins. As this method is costly and requires expertise, this study focused on visualizing NET using light microscopy. Method: Neutrophils were isolated by density gradient centrifugation. The cells were adhered on a gelatin-coated coverslip and stimulated using S. aureus cell-free culture supernatant for 10 or 30 minutes for N(ET) release. The slides were stained with a combination of Hoechst and Giemsa stains and viewed under the light microscope. Result: NET-DNA was observed more clearly and prominently by this double staining method using a light microscope. Conclusion: This method helped in seeing NET-DNA with more resolution under the light microscope. Since the dyes used for staining are cost effective, this method can be employed for preliminary NET research in labs with limited resources.
PAGES: 221-225  |  20 VIEWS  31 DOWNLOADS

How To Cite this Article:

Haritha V H, Binchu V Shaji and *Anie Y. Visualization of Extracellular Trap by Light Microscopy. 2019; 9(1): 221-225.