测定样品的荧光激发和发射最大值

智能文字提取功能测试中

进行荧光测量的挑战之一是确定新样品的最佳激发和发射波长。由于使用荧光技术可以获得大量信息，因此优化测量参数以获得尽可能好的数据至关重要。Jasco开发了一个三维荧光测量程序，该程序将搜索用于未知样品的最佳峰值激发和发射波长。这是通过在改变激发波长的同时测量样品的发射光谱来实现的，或者相反，通过在发射波长改变时测量激发光谱来实现。本应用说明演示了如何确定样品的激发和发射最大值，以获得使用三维荧光测量程序的更深入的荧光测量。Application Note 2/3 Determining a Sample's Fluorescence Excitation and Emission Maxima Introdu c tio n One of t h e c h al l enges i n performing f l uorescence measurements i s determi n ing t h e opti m um excitatio n and emission wavelengths for a new sample. With the vast amount of in f ormat i on tha t ca n be obtained using fl u orescence techniques， it is cr i tical to optimize the measurement parameters to obtain the best poss i ble data.Jasco has developed a 3-D f lu orescence measurement program that will search for the optimum peak excitat i on a n d em i ssion wavelengths to u se for u n known samples. This is accomplished by measur i n g the emission spectrum of a sample while changing it s exc i tation wavelengt h or， conversely， by me a surin g t h e excita ti on spect ru m as th e em i ssion wave l ength is c h anged. This appl i cation note shows the 3-D f luorescence spectra of p-terphenyl and tonic water us i ng a FP-6500. Keywo r ds FP-6500， Fluorescence Exp e rimental Data Acquisition Interval 1nm Response Time 4 seconds Spectral Bandwidth 1nm Scan Speed 20 nm/min Determin i ng a Sample's Fluorescence Appl i cat i on Note Excitation and Emission Maxima R esul t s The excitatio n and emiss i on spect r a fo r p -terphenyl and t onic water we r e o b tai n ed u s i n g an FP-6500 spectrof l u r o m ete r and a 10 m m pa th length cell . Figu r e 1 illustrates the 3-D plot of t h e fl u orescence intensity of p-terphenyl as a f unct i on of excitation and emission wavelengths. T h e maximum exci t a ti on wave l ength occurs at 295 nm while the maximum emiss i on wavele n gt h i s at 340 nm. Figure 1. 3-D gra ph i ca l r eprese n t a tio n o f t he exc i tat i o n a nd emi s s i on s p ect r a of p -Ter p henyl. Qu in i n e i s a st r o n gly fl u or e sc en t com p o u nd that i s found in tonic wat e r . I n dil u t e acidic sol u tions， t h er e ar e two abso r ptior bands ce n ter e d at 250 a n d 350 nm， whi l e t h e maximum flu orescence ba n d occurs at 450 n m. T h e 3-D f l uo r escence spec t rum of t on i c water (Figure 2) was measured a n d then analyzed using t h e I nterval Data Analysis program . The maximum excita ti o n a n d emission wave l engths d e te r min e d by th e prog r am were 360 and 455 nm， r e s p ectiv e ly. Figure 2. 3-D grap h ical r epresentat i on of the excitation an d e m ission spectra of tonic water. 28600 Mary's Cour t ， E a s to n ， M D 21601US A UAGE O A ppli c a tio n Lib r ary： htt p：//www.jas co in c .c o m/a pp l icatio n s Figure 3. Emissi o n s p ect r um of tonic water . C onclusion This application n ote demonstrates how to deter m ine a sample's excitation and emissio n maxima for obtaini n g more i n -depth fluorescence measurements using the 3-D F lu orescence Measu r eme n t Program. 28600 Mary's Cour t ， E a s to n ， M D 21601US A