Hitachi F 2500 Manual

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(2) Operating Procedures 
 
(a)  Set PMT Voltage to 400 V and Data Mode to 
“Fluorescence”.    Perform “Zero Adjust” to calibrate 
the zero point.     
 
(b)  Set the triangular cell filled with Rhodamine B into the 
cell holder in the sample compartment.    (See Fig. 
F-2).   
The triangular cell must be set in the direction shown 
in Fig. F-3. 
 
 
 
 
 
 
 
 
 
 
 
 
Fig. F-2    Setting of Rhodamine B 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Fig. F-3    Setting Direction of Triangular Cell (Top view)...

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(c)  Select “EX” of “Instrumental Response:    Utility”. 
EM wavelength is set at 640 nm and the instrument 
starts EX wavelength scan (from 198.6 nm to   
601.4 nm).   
At this stage, the instrument is automatically set in the 
following conditions.   
 
Slit (EX/EM) ........... 5.0/20.0 nm 
Scan speed ............ 60 nm/min 
Scan mode............. Fluorescence 
PMT Voltage .......... 400 V 
Response............... Auto 
 
(b)  Upon completing the measurement, all of measuring 
conditions are...

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F.2    Measurement of Instrumental Response on Emission Side 
 
This is the function to obtain the instrumental response on the 
emission side such as wavelength characteristics of the emission 
monochromator and detector (photomultiplier).     
The instrumental response on the emission side is determined by 
measuring a combination of instrumental response on both the 
excitation and emission sides by synchronous wavelength scan, 
and dividing it by the instrumental response on the excitation...

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(c)  Select “EM (200 to 600 nm)” of “Instrumental 
Response:  Utility”.   
EM wavelength is set at 198.6 nm and the instrument 
starts synchronous wavelength scan (from 198.6 nm 
to 601.4 nm).     
At this stage, the instrument is automatically set in the 
following conditions.   
 
Slit (EX/EM) ......... 5.0/20.0 nm 
Scan speed .......... 60 nm/min 
Scan mode .......... Fluorescence 
PMT Voltage ........ 400 V 
Response ............ Auto 
 
(d)  Upon completing the measurement, all of measuring...

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F.3  Measurement of Instrumental Response on the Long Wavelength Range 
of Emission Side (only with Model F-4500) 
 
When a corrected spectrum in the long wavelength area of   
600 nm is required on the emission side, the beam of a lamp for 
which the wavelength characteristics are known is allowed to be 
irradiated to the emission optics, and the instrumental response 
in the long wavelength range on the emission side is read for the 
correction.   
As a light source, a sub standard light source...

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(e)  Upon completing the measurement, all of measuring 
conditions are set to those set before.     
Now, instrumental response on the emission side has 
been measured and stored.    Remove the sub 
standard light source.     
  

 INDEX - 
1 
INDEX 
 
 
A 
ASCII text file  ............................................................................................................. 1-3, 6-25 
Activity ....................................................................................................................... 3-16, A-6 
Analog output gain  ................................................................................................... 1-41, 8-10 
Auto pre-scan...

 INDEX - 
2 
F 
Factor input  ......................................................................................................................... 4-7 
Fonts tab  ........................................................................................................................... 1-32 
 
 
G 
General tab  .................................................................................................... 2-3, 3-2, 4-2, 5-3 
Gradient...

 INDEX - 
3 
N 
Number of wavelengths ...................................................................................................... 4-10 
 
 
P 
Peak area  ........................................................................................................................... 4-5 
Peak detection  ......................................................................................... 1-3, 2-12, 3-10, 5-10 
Peak height...

 INDEX - 
4 
Spectral calculation  ........................................................................................................... 6-16 
Spectrophotometer menu  ................................................................................................. 1-40 
Spectrum toolbar  ..................................................................................................... 1-20, 1-22 
Stack...