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FL 6500/8500 Integrating Sphere Installation
Instructions
This instruction shee t describes the installation of this a ccessory which is used with the FL 6500/8500
Fluorescence Spectrometer.
NOTE:
Read these instructions before you install this accessory.
Contacting PerkinElmer
Supplies, replacement parts, and accessories can be ordered directly from PerkinElmer, using the part
numbers.
See our website:
http://perkinelmer.com
PerkinElmer's catalog service offers a full selection of high-quality supplies.
To place an order for supplies and many replacement parts, request a free catalog, or ask for
information:
If you are located within the U.S., call toll free 1-800-762-4000, 8 a.m. to 8 p.m. EST. Your order will
be shipped promptly, usually within 24 hours.
If you are located outside of the U.S., call your l oca l PerkinElmer sales or service office.
Features
• Used for getting Quantum Yield
• Available sample type: Powder/Liquid
• Provide the Simplification and the de Mello’s method
Figure 1 FL 6500/8500 Integrating Sphere [P/N:N4201017]
PerkinElmer, 710 Bridgeport Avenue,
Shelton, CT 06484-4794, U.S.A
Produced in the USA.
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Dimensions and Specifications
Physical characteristic Specification
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Outline
Weight
Integrating Sphe re
Sample Holder size
Dimensions (mm) 130 x 267 x 170 (WDH)
Kg 2.86
Diameter (mm) 101.6
mm 12.5 x 12.5
Connectable Cells
Description
Standard cell, 10 x 10 mm
Micro cell, 10 x 10 mm
Configuration of the Integratin g Sphe re
Figure 2 Integrating Sphe re Configuration
Figure 3 Quartz Cell with Low Profile Stopper [P/N: N4202030]
NOTE:
When using the integrating Sphere, it is recommended to use the enclosed Quartz Cell to
prevent scratching of the lid.
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Figure 4 Lid an d Cel l Ho lder Cap
Installation
1. Prepare the FL 6500/8500 Fluorescence Spectrometer to insta ll this accessory.
2. Connect the power cord and the communication cable.
3. Loosen the accessory fixing bolt a nd remove the current sample accessory.
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Figure 5 Loosening the Accessory Fixing Bolt
4. Pull out the cell holder by hand.
Figure 6 Pulling Out the Cell Holder
5. Prepare an Integrating Sphere. Make sure that there is no sample inside before mounting the
Integrating Sphere.
Figure 7 Checking the Sample Holder
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6. After checking the pogo pin position of the sam ple compartment, place t he Integrating Sphere
into the pogo pin well.
Figure 8 Install the Accessory
7. Tighten the accessory fixing bolt.
Figure 9 Tightening the Accessory Fixing Bolt
8. Remove the lid part of the Integrating Sphere and separate the cell holder cap.
Figure 10 Remove the Lid Part
9. After loading the sample cell into the separated cell holder cap, mount it in the accessory and
measure it.
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Correction Spec tra for Integrating Sphere
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NOTE:
NOTE:
When using the integrating Sphere for the first time, a correction factor for the integrating
sphere must be created and stored.
The spectrum could be altered because of light scattering, or instrumental condition like lamp
fluctuation. It could be correct ed by Rhodamine 101 and MgO diffuser as reference material.
The correction factor is then produced and saved. This factor is various among instruments
(because of influence of the devices such as la mp, detector, and so on) , so it can’t be used
with other instrument even in the same model or maker. Spectra function is used to
compensate the spectra’s distortion which may be caused by the differences in reflectance,
absorptivity, sensitivity of each component of the system such as PMT, grating , mirror, lens,
etc.
Correction tools (Rhodamine 101 cell, MgO diffuser) are optional accessory.
To correct sp ectra, Rhodamine 101 is required as a separate purchased. The MgO diffuser is
prepared by user. Prepare a solution of about 2 mg MgO in 10 mL distilled water.
1. How to measure the Excitation and Emission correction factors
1. Execute the FLServiceTool software.
2. Select the Calibration tab in the Mode, click Spectrum Correction.
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3. After setting parameters as below picture, click the Max RFU icon. When the Max RFU
measurement is finished, click the Start button.
4. When the pop-up wi ndow appears, place the Rhodamine 101 triangular cell in the integrating
sphere, Click Yes to start measurement.
NOTE:
The mounting direction of Rhodamine 101 is as follows.
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5. After the measurement is finished, select the measured graph and click the Save Correction
button.
6. Click Yes to the Ex Correction factor for Integrating Sphere.
NOTE:
Be careful not to save the file name as an existing EX.cor.
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7. After saving the excitation correction factor for Integrating Sphere, select Synchronous
(Low) in the Scan Mode of the Spectrum Correction Settings tab.
8. Click Start. When pop up window appears, click OK.
9. Select the saved excitation factor for the integrating sphere and click Open.
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10. When the pop up window appears, afte r shaking t he prepared MgO diffuser well and mount it.
Click Yes.
11. When the measurement is completed from 200 to 600 nm, the following pop-up window will
appear. Click OK and select the saved EM. cor File in the existing device.
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12. Confirm that data of 600 ~ 850 nm is added.
13. Select the measured Synchronous spectrum and click Save Correction button.
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14. Click Yes to save the Emission Correction factor for Integrating Sphere.
NOTE:
Be careful not to save the file name as an existing EM.cor.
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2. How to apply the correction factor in measurement
1. Execute the Spectrum FL software.
2. Select the Quantum Yield mode.
3. When setting parameters, select File in the Ex.corr tab.
4. Click the File select button and select the saved correction factor file. Click OK.
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5. Apply the correction factor in the same way as in steps 3-4 for the Emission.
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Measurement
Perform the measurements according to the purpose of the experiment.
• Absolute mode: Simplificati on method
Quantum yield = Ec/(La-Lc)
1. Double click on the Spectrum FL software.
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2. Check the recognition of Accessory.
3. Click Quantum Yield mode.
4. Select Absolute Calc Mode and Simplification Calc Method in the Data Collection tab.
5. Set up the measurement parameters in turn.
NOTE:
In Absolute mode, select File from Ex. Corr and Em. Corr tab, then select the corre ction factor
for integrating sphere. Refer to the
on the page 10.
6. Click Save to save the method after setting up the parameters.
7. Select the Run icon.
8. Input the sample name and click OK.
9. Input Solvent or Empty cell. And click OK.
10. R emove Solvent or empty cell and input the Samp le (Direct position) click OK.
11. Confirm the spectrum and results.
12. Calculate the quantum yield in the result window.
2. How to apply the corr ectio n fa ctor in measurement
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13. Enlarge the graph inter section.
14. Check the intersection value and separates the excitation and emission areas of each graph
based on the intersection value.
15. When the area is set, the Quantum yield value is automatically calculated and displayed.
16. Save or print the data.
• Absolute mode : De Mello method
Quantum yield = Ec-[(1-A)*Eb]/La*A
A = 1-(Lc/Lb)
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1. Double click on the Spectrum FL software and select the measurement mode.
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2. Check the recognition of Accessory.
3. Click the Quantum Yield mode.
4. Select Absolute Calc Mode and De Mello Calc Method in the Data Collection tab.
5. Set up the measurement parameters.
NOTE:
For more detail of method, refer to Spectrum FL Software Users Guide.
6. Click Save to save the method after setting up the parameters.
7. Select the Run icon.
8. Input the sample name and select OK.
9. Input Solvent or Empty cell. And select OK.
10. Remove Solvent or empty cell and input the Sample.
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11. Turn the knob to Indirect position, select OK.
12. Turn the Knob to Direct position, select OK.
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13. Confirm the spectrum and results.
14. Calculate the quantum yield in the result window.
15. Enlarge the graph intersection.
16. Check the intersection value and separates the excitation and emission areas of each graph
based on the intersection value.
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17. When the area is set, the Quantum yield value is automatically calculated and displayed.
• Relative Quantum Yi eld
Relative Quantum Yield method is not required an Integrating Sphere accessory. This method is
determined by comparing the reference with known quantum yield with the sample in question.
And it is needed Absorbance and Refractive Index values.
1. Double click on the Spectrum FL software and select the measurement mode.
2. Check the recognition of Accessory.
3. Click the Quantum Yield mode.
4. Select the Relative Calc Mode in the Da ta Collection tab.
NOTE:
Standard and sample are measured and compared under the same conditions. Therefore, errors
can be minimized by using substances with simila r emission ranges.
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5. Set up the measurement parameters.
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NOTE:
NOTE:
In Relative Quantum Yield mode, select On from Ex. Corr and Em. Corr tab.
6. Select sample table Tab and add the sample.
One of the samples must be set to standard, and the Absorbance, Refractive and Quantum Yield
values must be entered.
7. Click Save to save the method after setting up the parameters.
8. Select the Run icon.
9. Input the sample name and click OK.
10. Input Standard sample cell. And click OK.
11. Remove Standard sample cell and input the Sample, click OK.
12. Confirm the spectrum and r esults.
13. Calculate the quantum yield in the result window.
14. Save or print the data.
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