Microfluidics emerged during the early 1990s with channel networks in silicon or glass. Microprocessing of these materials is labor-intensive and time-consuming, it requires sophisticated equipment in a clean room, and often involves hazardous chemicals. The subsequent use of polymer greatly simplified the fabrication of microchips and led to the rapid development of the field. Polymer such as poly(dimethylsiloxane) (PDMS), has other attractive properties, such as being elastic (easy to make efficient microvalves), permeable to gases, and compatible with culturing biological cells. Despite these advantages, applications of PDMS chips are severely limited by a few drawbacks that are inherent to this material: (i) strong adsorption of molecules, particularly large biomolecules, onto its surface; (ii) absorption of nonpolar and weakly polar molecules into PDMS bulk; (iii) leaching of small molecules from PDMS bulk into solutions; and (iv) incompatibility with organic solvents. To overcome all these problems, Teflon plastics seem to be the perfect solution. They are well-known for their superior inertness to almost all chemicals and all solvents; they also show excellent resistance to molecular adsorption and molecule leaching from the polymer bulk to solutions. However, Teflon has a high chemical inertness of the surface, which is restricted the bonding temperature (>260¢XC).It is not conducive to the low-temperature packaging process. This study presents a simple and rapid process for sealing Teflon-based microfluidic chip at a temperature of 140oC which is lower than typical bonding temperature of 260oC. A simple ammonium plasma treatment is used to enhance the surface energy of Teflon substrates such that the bonding temperature can be greatly reduced. Results indicate that the ammonium plasma treated Teflon substrates can be sealed using hot press bonding at a temperature of 140oC for 20 min. The measured iv bonding strength for the Teflon-based microfluidic devices is higher than those bonded at a reported temperature of 260oC for 60 min. It shows the measured contact angle for the Teflon substrates treated with different plasmas. Results indicated that the ammonium hydroxide plasma exhibited the best wettability property and the contact angle reached the minimum value of 45o after 5 min of treatment. The ESCA analysis showed the best Defluorination by ammonium plasma. The fluorine/carbon atomic ratio degraded from 1.96 to 1.10 by 5 minutes. The measured bonding strength for the Teflon substrates bonded with different surface activation protocols. Results showed that the bonding strength was enhanced upto 93% after the plasma treatment. The plasma treatment not only enhanced the bonding strength but also reduced the bonding temperature and time. The measured surface roughness only increased 15¡Ó5 nm (Ra) after the plasma treatment, which is acceptable for most applications in microfluidic systems. Finally, the fluorescence optical architecture and cross-chip successfully detected and isolated £XX-174 fragment of DNA samples confirmed the Teflon substrate for the emerging microfluidic plastic chip. The developed method provides a simple and rapid way to fabricate Teflon-based microfluidic devices.