PerkinElmer FL 6500 User manual

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PerkinElmer, 710 Bridgeport Avenue,
Shelton, CT 06484-4794, U.S.A
Produced in the USA.
FL 6500/8500 Microplate Reader Installation Instructions
This instruction sheet describes the installation of this accessory 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 local PerkinElmer sales or service office.
Features
• Plug-n-play accessory
• Measure samples in any plate format (Default: 96 or 384 well)
• Create new plate formats graphically and interactively
Figure 1 Microplate Reader and cover (P/N: N4201024)

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Dimensions and Specifications
Physical Characteristic Specifications
Microplate Reader Dimensions (mm) 218(W) x 340(D) x 290(H)
Microplate Reader Weight (Kg) 4.2
Microplate Reader Cover (mm) 259(W) x 430.5(D) x 331(H)
Microplate Reader Cover Weight (Kg) 4.25
Available Microplates
Physical Characteristic Specifications
The number of Wells Default: 96 or 384 and customize available
Microplate Size (mm) 85.4 x 127.6
Configuration of the Microplate Reader
• Microplate Reader
Figure 2Microplate Reader
a. Microplate fixing jig
b. X and Y axis stage
c. Microplate Reader fixing bolt
d. Collimator Lens Holder (Emission)

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e. Collimator Lens Holder (Excitation)
f. Collimator Lens Holder (Sample)
g. Optical Fiber
h. M1 Cable
i. M2 Cable
j. Pogo Pin Bracket
• Cover for Micorplate Reader
Figure 3Microplate Reader Cover
• Two cradles for the Microplate Reader
Figure 4Cradles
Installation
1. Prepare the FL 6500/8500 Fluorescence Spectrometer to install this accessory.
2. Connect the instrument power cord and the communication cable.
3. Loosen the accessory fixing bolt and remove the existing sample accessory.
Figure 5Loosening the accessory fixing bolt

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4. Pull out the cell holder by hand.
Figure 6Pulling out the cell holder
5. Remove the existing front cover of the FL 6500/8500 using a screwdriver.
Figure 7 Front Cover
6. Place the front cover of Microplate Reader and fasten the three bolts using a screwdriver.
Figure 8 Front Cover of Microplate Reader
7. Undo the four screws which attach the Hinge Guide to the sample compartment lid.
Figure 9 Remove the Hinge Guide

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8. Undo the four screws which attach the sample compartment lid to the FL 6500/8500.
Figure 10 Remove the sample compartment lid
9. Tighten the four screws that secure the cover of the plate reader to the FL 6500/8500.
Figure 11 Cover for the Microplate Reader
10. Attach the Microplate Reader cradles to the sample compartment.
Figure 12 Cradles of the Microplate Reader

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11. Fix the M4 bolts under the cradles using a 3 mm wrench.
Figure 13 Fix the M4 bolts
12. Connect the M1 and M2 cables of the Microplate Reader into the accessory port.
Figure 14 Connect M1 and M2 cables
13. Fit the groove (hole) of the Microplate Reader into the position of the cradles. (If positioned well,
the pogo pin will fit also.)
Figure 15 Install the Microplate Reader

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14. Tighten the accessory fixing bolt.
Figure 16 Accessory fixing bolt of Microplate Reader
15. Close the Microplate Reader cover and then, lock the latch at the left and right side of the front
cover.
Figure 17 Lock the latches
NOTE:
Sample loading can be done by opening the upper door of the Microplate Reader.
Figure 18 Upper door of the Microplate Reader

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Calibration of Microplate Reader
1. Execute the Spectrum FL software.
2. Select Options mode →Microplate Reader icon Or click the
Accessory icon .
3. The following window will appear. Select the plate you want to use.
4. Enter the required parameters on each tab if you know the whole plate information.
List Function
Plate Setting
Select the type of well plate. Default is 96 or 384 wells. Customized
setting is available.
Column Count Enter the value for the number of columns.

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Row Count Enter the value for the number of rows.
Start X, Start Y
Enter the x and y distance of the A1 well to the upper right corner of
the plate.
Interval X,
Interval Y
Enter the distance between the centers of two neighboring wells in
the x and y directions.
Radius (mm) Enter the value for the radius of the wells in the plate.
Reload Open the previously saved parameters.
Save Save the setting parameters.
Manual Alignment Refer to the Manual Alignment chapter.
Show Dimensions
If you check Show Dimensions, it shows the range of each
parameter.
5. Click Save.
6. Click the Lamp On button in the Operation to turn on the lamp. Enter the A1 and click Move
to button.
7. If the A1 position is not correct, Go to the next Manual Alignment section.
NOTE:
The Start X value and the Start Y value may differ because the starting position differs depending on
the accessories and equipment.

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Manual Alignment
NOTE: If you do not know the Plate information or A1 position is not correct, proceed the manual alignment.
1. Prepare a micro plate and Scotch®MagicTM Tape. Attach the magic tape onto the A1 and H12
(96 well) or P24 (384 well) area as shown below.
2. Load the micro plate.
3. Click Manual Alignment in the Plate Information.

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4. The following window appears.
List Function
Movement Setting Use these settings to define the distance the probe is moved head, each time an alignment arrow is pressed. Use 10 mm for
the coarse tuning and 0.1 mm for the fine tuning.
Movement Control
Use the arrows to align the probe. Each time you click on one of
the arrows the probe is moved in the indicated direction. The
distance the probe is moved is determined by the alignment
distance.
Operation
List Function
Move to Start Position Move the micro plate to the start position.
Turn on Lamp Turn on the lamp.
Turn off Lamp Turn off the lamp.
5. Enter the Row and Column counts in the Step 1. Then click Done.
6. The lamp will be turned on automatically.

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7. Click Done in Step 2.
8. Align the beam to the centre of A1 using the Movement Control icons and click Done.
9. Align the beam to the centre of H12 using the Movement Control icons and click Done.

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10. Align the beam to the edge of H12 using the Movement Control icons and click Done.
11. Click Finish to save the plate information and close the window.
12. You can see that the Plate Information tab has been changed.

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13. Select any position in the micro plate picture and click Save and then Move to. Confirm that the
beam is aligned well.
14. If the beam position is correct, Go to the following Measurement section. If the beam position is
not correct, Execute the Manual Alignment again.
Measurement
1. Open the Spectrum FL software and select a Microplate Reader mode in the Method Setup.
2. In the Data Collection tab, set the measurement parameters in turn.
NOTE:
For more detail of method, refer to Spectrum FL Software Users Guide.
3. When you click the Excitation of the Instrument Setting, enter the number of the wavelength for
measurement in the Measured Num. The ID in the Excitation Setup is generated as entered in
Measured Num. Enter the wavelengths and slits for the experiment.

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4. When you click the Accessory of the Instrument Setting, the following window appears.
a. When you select Center of Cell, the following setup window appears.
List Function
Center of Cell Selecting this means that whenever you click on a well to set a measurement point, the measurement point is automatically set
to the center of the well.
Cycle Number This is the number of measurement cycle.
Time Interval
The time interval is the time between last measurement of current
cycle and the 1st measurement of next cycle.
Before the next cycle measurement, a message window will pop
up as shown below. Click Skip to measure immediately.
b. When you select Customize, the following setup window appears.

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List Function
Customize
Selecting this means that whenever you click on a well to set a
measurement point, the measurement point is set exactly where you
clicked, rather than being automatically centered on the well.
Normally the customize option is used for a large well (eg. 4 or 16)
Calculation Mode
Select Average or Sum.
Average: If selected, multiple reads for a given well are averaged.
Sum: If selected, multiple reads for a given well are summed
(integrated). The results spreadsheet is updated when all readings
are performed for a given well.
Cycle Number This is the number of measurement cycle.
Time Interval
The time interval is the time between last measurement of the
current cycle and the first measurement of next cycle.
Before the next cycle measurement, a message window will pop up
as shown below. Click Skip to measure immediately.
5. After setting the parameters in the Data Collection, select Sample Tables tab. Add the sample
numbers and select the each sample type.

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6. Click the Position to select the cell position.
7. The following window will appear. Select the cell position using a mouse.
NOTE:
Each selection: Click the mouse, Each cancellation: Click the selected position again.
Area selection: Drag the mouse, Area cancellation: Click the mouse outside the plate anywhere.
8. Click Save to save the method after setting up the parameters.
9. Prepare a Microplate filled with the sample for measurement.
10. Load the Microplate.
11. Select the Run icon.
12. Input the sample name and select OK.
13. Save or print the data if required.

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