Hitachi F7000 Instruction Manual

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(6)  As shown in Fig. F-2, set the triangular cell filled with Rhodamine 
B into the cell holder in the sample compartment, the filter holder 
to the sample compartment and the red filter (single-side frosted) 
in the filter holder.    In so doing, pay attention to the direction of 
the triangular cell and filter.    (Refer to Fig. F-3) 
 
 
 
 
Fig. F-2    Setting of Triangular Cell Containing Rhodamine B 
 
 
 
 
Fig. F-3    Setting Direction of Triangular Cell (top view) 
 
 
NOTE:  It may take time to obtain stable data due to characteristics of 
the detector immediately after the photomultiplier voltage has 
been remarkably changed (from 700 V to 250 V for example).   
When more accurately calibrating the zero point, wait until 
stable data is obtained and then proceed to step (5). 
  Triangular cell with 
fixture (containing 
Rhodamine B) 
Filter holde
rRed filter 
Metal part
Excitation light
Triangular cellFluorescenceRed filter   
(single-side frosted)
Frosted  
						

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(7)  Execute the Instrumental Response command from the Utility 
menu.    The Instrumental Response dialog box will open as 
shown in Fig. F-4. 
Select “EX [200 to 600 nm].”    (Refer to ①  in Fig. F-4.) 
 
 
 
Fig. F-4 
 
 
(8) Click the Run button.  (Refer to ②  in Fig. F-4.) 
A message box “Set the triangular cell with Rhodamine B to cell 
holder and set the red filter.” will appear.    After setting the 
Rhodamine B and red filter, click the OK button.  (Refer to ③ in 
Fig. F-5.) 
 
 
 
Fig. F-5 
 
 
1 
2
3 
						

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(9)  The EX wavelength is set at 640 nm and the instrument starts EX 
wavelength scan (from 198.6 to 601.4 nm). 
At this stage, the following measuring conditions are automatically 
set. 
• Scan mode :  Excitation 
• Data mode :  Fluorescence 
•  EX slit  :  5.0 nm 
•  EM slit  :  20.0 nm 
•  Scan speed  :  60 nm/min 
•  PMT voltage :  400 V 
• Response  :  Auto 
 
(10)  Upon completion of measurement, the conditions set before 
measurement are retrieved. 
Now, the instrument response on the excitation side has been 
measured and stored.    Take out the rectangular cell. 
  
						

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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 
or 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 side 
preliminarily measured.    An emission spectrum is correctable within a 
range of 200 to 600 nm. 
 
 
(1) Click the 
  (analysis method) button on the Measurement 
toolbar.    A box for setting your analysis method will appear. 
 
(2)  Select the General tab.    On the General tab page, specify 
“Wavelength scan” for the measurement mode. 
 
(3)  Select the Instrument tab. 
 
(4)  Set “Fluorescence” for the data mode and “250 V” for the 
photomultiplier voltage. 
 
(5)  Execute the Zero Adjust command from the Spectrophotometer 
menu to calibrate the zero point. 
 
(6)  As shown in Fig. F-6, set the diffuser into the cell holder in the 
sample compartment.   
 
 
 
Fig. F-6    Setting of Diffuser 
F.2.1 Overview 
F.2.2 Operating 
Procedures 
 
Diffuser  
						

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NOTES:  1.  Measurement of the instrumental response on the 
excitation side must be completed before executing 
measurement of the instrumental response on the 
emission side. 
  2.  It may take time to obtain stable data due to 
characteristics of the detector immediately after the 
photomultiplier voltage has been remarkably changed 
(from 700 V to 250 V for example).    When more 
accurately calibrating the zero point, wait until stable 
data is obtained and then proceed to step (5). 
 
(7)  Execute the Instrumental Response command from the Utility 
menu.    The Instrumental Response dialog box will open as 
shown in Fig. F-7. 
Select “EM [200 to 600 nm].”    (Refer to ①  in Fig. F-7.) 
 
 
 
Fig. F-7 
 
 
(8) Click the Run button.  (Refer to ② in Fig. F-7.)  A message 
box “Set diffuser to cell holder.” will appear.    Click the OK button.   
(Refer to ③  in Fig. F-8.) 
 
 
 
Fig. F-8 
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2
3 
						

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(9)  The EX and EM wavelengths are set at 198.6 nm and the 
instrument starts synchronous wavelength scan (from 198.6 to 
601.4 nm). 
At this stage, the following measuring conditions are automatically 
set. 
• Scan mode :  Synchronous 
• Data mode :  Fluorescence 
•  EX slit  :  5.0 nm 
•  EM slit  :  20.0 nm 
•  Scan speed  :  60 nm/min 
•  PMT voltage :  250 V 
• Response  :  Auto 
 
(10)  Upon completion of measurement, the conditions set before 
measurement are retrieved. 
Now, the instrument response on the excitation side has been 
measured and stored.    Take out the diffuser. 
 
  
						

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F.3  Measurement of Instrumental Response in 
Long Wavelength Range on Emission Side 
 
When a corrected spectrum in the long wavelength area of 600 nm or 
more is required on the emission side, the beam of a lamp whose 
wavelength characteristics are known is allowed to be irradiated to the 
emission monochromator, and the instrumental response in the long 
wavelength range on the emission side is read for correction. 
As a light source, a sub standard light source (P/N 250-0123) is used.   
The correction range by it is 500 to 900 nm and, when combined with an 
instrumental response in the short wavelength range, spectrum 
correction is available over the entire wavelength range (200 to 900 nm) 
on the emission side. 
 
 
 
Refer to the instruction manual for the sub standard light source 
(optional accessory). 
 
(1)  Close the cover of the sample compartment. 
 
(2) Click the 
  (analysis method) button on the Measurement 
toolbar.    A box for setting your analysis method will appear. 
 
(3)  Select the General tab.    On the General tab page, specify 
“Wavelength scan” for the measurement mode. 
 
(4)  Select the Instrument tab. 
 
(5)  Set “Fluorescence” for the data mode and “400 V” for the 
photomultiplier voltage. 
 
(6)  Execute the Zero Adjust command from the Spectrophotometer 
menu to calibrate the zero point. 
 
(7)  Set the sub standard light source in the sample compartment with 
reference to its instruction manual.    (Refer to Fig. F-9.) 
Turn on the power switch of the sub standard light source and 
wait for approx. 5 minutes until the lamp stabilizes. 
F.3.1 Overview 
 
F.3.2 Operating 
Procedures  
						

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Fig. F-9 
 
 
(8)  Execute the Instrumental Response command from the Utility 
menu.    The Instrumental Response dialog box will open as 
shown in Fig. F-10. 
Select “EM [500 to 900 nm].”    (Refer to ①  in Fig. F-10.) 
 
 
 
Fig. F-10 
 
 
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2 
						

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(9) Click the Run button.  (Refer to ② in Fig. F-10.)  A message 
box “Set the sub standard light source.” will appear.    Click the 
OK button.  (Refer to ③  in Fig. F-11.) 
 
 
 
Fig. F-11 
 
 
(10)  The instrument starts EM wavelength scan (from 498.6 to   
901.4 nm). 
At this stage, the following measuring conditions are automatically 
set. 
• Scan mode :  Emission 
• Data mode :  Luminescence 
•  EM slit  :  5.0 nm 
•  Scan speed  :  60 nm/min 
•  PMT voltage :  400 V 
• Response  :  Auto 
 
(11)  Upon completion of measurement, the conditions set before 
measurement are retrieved. 
Now, the instrument response on the emission side has been 
measured and stored.    Take out the sub standard light source 
and diffuser. 
 
 
3 
						

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F.4  Measurement of Instrumental Response in 
Long Wavelength Range on Excitation Side 
 
When a corrected spectrum in the long wavelength area of 600 nm or 
more is required on the excitation side, the instrumental response in the 
long wavelength range on the excitation side is read for the correction 
with reference to the instrumental response obtained at the long 
wavelength on the emission side. 
This measurement is enabled only when the instrumental response at 
the long wavelength on the emission side is available.    The correction 
range by it is 500 to 900 nm and, when combined with an instrumental 
response in the short wavelength range, spectrum correction is 
available over the entire wavelength range (200 to 900 nm) on the 
excitation side. 
 
 
(1)  Close the cover of the sample compartment. 
 
(2) Click the 
  (analysis method) button on the Measurement 
toolbar.    A box for setting your analysis method will appear. 
 
(3)  Select the General tab.    On the General tab page, specify 
“Wavelength scan” for the measurement mode. 
 
(4)  Select the Instrument tab. 
 
(5)  Set “Fluorescence” for the data mode and “350 V” for the 
photomultiplier voltage. 
 
(6)  Execute the Zero Adjust command from the Spectrophotometer 
menu to calibrate the zero point. 
 
F.4.1 Overview 
 
F.4.2 Operating 
Procedures