[±×¸²3. Rapid mixing accessoryÀÇ ¸ð½Äµµ]

1. Ãʱâ NATAÀÇ Çü±¤ ¼¼±â¸¦ ÃøÁ¤Çϱâ À§Çؼ­ Syringe A¿¡ 5 uM NATA¸¦ ÁÖÀÔÇϰí, Syringe B ¿¡ ¿ë¸ÅÀÎ 0.1 M Sodium phosphate¸¦ ÁÖÀÔÇÑ´Ù.
2. Çü±¤ cell¿¡ ÁÖÀ﵃ ½Ã·áÀÇ ¾çÀ» ¾à 500 uLÁ¤µµ·Î ¼³Á¤ÇÑ ÈÄ Push bar¸¦ ´­·¯ ¼¿¿¡ ½Ã·á¸¦ ÁÖÀÔÇÑ´Ù. ¼³Á¤µÈ ¾ç ¸¸Å­ÀÇ ½Ã·á°¡ ÁÖÀԵǸé stop syringe°¡ trigger ½ÅÈ£¸¦ ÁÖ°í Çü±¤°è°¡ Çü±¤ ÃøÁ¤À» ½ÃÀÛ ÇÑ´Ù. À§ °úÁ¤À» 6ȸ ¹Ýº¹ÇÑ´Ù.
3. ÃÖÃÊ 1ȸ ÃøÁ¤ÇÑ µ¥ÀÌÅ͸¦ Á¦¿ÜÇÑ 5°³ÀÇ µ¥ÀÌÅ͸¦ Æò±ÕÇÑ´Ù. ÀÌ µ¥ÀÌÅÍ´Â Ãʱâ NATAÀÇ Çü±¤ ¼¼±â °ªÀÌ´Ù.
4. Syringe B¿¡ ÁÖÀÔµÈ 0.1 M Sodium phosphat- e¸¦ ¸ðµÎ ¹èÃâ½Ã۰í 50 uM NBS¸¦ ÁÖÀÔÇÑ´Ù. Push bar¸¦ ´­·¯ ½Ã·á¸¦ µ¿½Ã¿¡ ÁÖÀÔÇÑ´Ù. Çü±¤ cell¿¡ ¼³Á¤µÈ ¾çÀÇ ½Ã·á°¡ ÁÖÀԵǸé stop syringe°¡ trigger ½ÅÈ£¸¦ ÁÖ°í Çü±¤°è°¡ Çü±¤ ÃøÁ¤À» ½ÃÀÛÇÑ´Ù. ÀÌ °úÁ¤À» 6ȸ ÀÌ»ó ¹Ýº¹Çؼ­ ù µ¥ÀÌÅ͸¦ Á¦¿ÜÇÑ ÀÌÈÄÀÇ µ¥ÀÌÅ͸¦ Æò±ÕÇÑ´Ù.
5. ÃøÁ¤µÈ µ¥ÀÌÅ͸¦ fitting programÀ» ÀÌ¿ëÇØ¼­ single exponential decay fittingÇÑ´Ù.

Data Mode: Fluorescence   EX Slit: 10 nm
PMT Voltage : 700 V   EM Slit: 10 nm
Integration Time: 5 ms   EX Filter: Air
EX Wave: 390 nm   EX Filter: Air
EM Wave: 460 nm   (v) Short Life Time
Interval Time: 5 ms   Total Time: 5000 ms