The instrument has a range of operation from 0.000 to 4.000 OD, but the specifications for accuracy, linearity and reproducibility only go to 2.500 OD. The range allows the user to make measurements above 2.500 OD and roughly determine how much the sample needs to be diluted if a very accurate value is required. The accuracy of values over 2.500 OD is limited as described below. (Note that some cuvette based spectrophotometers are only specified for accuracy to 1.0 OD, but can read up to 5.0 OD.)
Microplate readers, like many other optical electronic instruments, have varying accuracy, repeatability, and linearity specifications that must become less stringent as higher OD values are approached. This is due to the nature of the reader’s optical and electronic sensing circuits. Assuming that a typical instrument is capable of parsing or dividing the dynamic signal range into exactly 60,000 “counts” then the following holds true:0 OD is 100% light (60,000 counts of signal),
Thus at very high ODs a variation of 0.1 OD is only a count of signal or less. This and other factors (such as bulb strength, ambient blocking etc.) do not allow the readers to always have the best results above a specific value. Samples that are very dark should be diluted in order to obtain the most accurate results.
Each laboratory must determine how accurate the results must be for the use of higher ODs. See FAQ “How can we verify that the measurements above the OD accuracy, linearity and repeatability specification are acceptable for our lab?”
The ELx800 and ELx808 microplate readers are compatible with parallel printers. Today, most printers are equipped only with USB ports. BioTek's readers cannot be directly linked to a printer USB port.
However, BioTek readers can be compatible to USB printers by using an LPT-USB adapter, such as the BioTek accessory p/n 75135. This adapter can also be purchased directly from the manufacturer, AK-Nord: http://www.ak-nord.de/ak/product_info.php?products_id=44 or other resellers.
There are several other HP printer models that can be compatible via this adapter. The key criteria is that the HP printer must support one of these HP printer languages:
* HP PCL3
* PCL 3e (enhanced)
* PCL 3 HPA
Because HP printer model numbers and availability vary by geographic region, it's best to confirm that the desired printer uses one of these three languages in order to be compatible with the HP printer output of the ELx800 and ELx808 readers.
Check that the following are used
1. 260 nm
2. UV transmissable plates with no fingerprints on the bottom
3. At least the minimum volume
4. Pathlength correction
5. 977 and 900 nm
6. Blank on the plate
7. Extinction coefficient of 50 is entered into the transformation spreadsheet
If the results are still not good enough, increase the volume to double the minimum
These instruments can be used for luminescence experiments if they are set up properly:
1. Turn off the instrument and remove the excitation filter wheel.
2. Replace one of the filters in the excitation wheel with a black plug (at least one should have come with your instrument).
3. Write down which position of the excitation filter wheel has the plug.
4. Return the excitation filter wheel to the instrument.
5. Remove the emission filter wheel.
6. Remove the filter in the emission filter wheel in position 1, position 4 is the second best choice (NOT position 2 or 3).
7. Write down which position of the emission filter wheel has the hole (empty position).
8. Return the emission filter wheel to the instrument.
9. Close the door and turn on the power on the instrument.
10. In Gen5 go to System/Reader Configuration, highlight your reader and click "view/modify," click "setup" then go to the "fluoresecence/luminescence" tab.
112. In the excitation wheel table, choose plug for the position with the plug.
12. In the emission wheel table, choose hole for the position with the hole.
13. Click "Send wavelengths."
14. In your Read step in the protocol, select Luminescence for the detection and the plug and hole will be chosen automatically.
15. The sensitivity for a luminescence experiment is typically in the range of 100-255. Sample readings should be > 5000 RLU.
Before calling or e-mailing to our service department (firstname.lastname@example.org), please collect the following information:
Serial number of the instrument. It is stamped onto a metal tag located either on the side or the back of most instruments (on the front inside door of the FLx800). The serial number will allow us to see exactly what instrument you have and with what options. We can also look up the instrument’s history, warranty and service contract information.
If you use a computer to drive the instrument: operating system , type and version of BioTek's software (can be found in the software menu, in the ''About...'' sub-menu).
Exact error codes you are experiencing and when they occur (upon startup, when you start a run, etc.).
a) To prevent damage to the reader or PC, always turn OFF the reader or the PC before removing or inserting a serial cable.
b) Reader controlling software and reader communication parameters will supersede the Windows settings. Windows serial port configuration settings should not need adjustment to enable proper communications. If you are using a USB cable or USB/serial adapter, you must install the proper USB driver. The USB connection will show up as a COM port in Windows Device Manager.
PC won’t communicate with reader:
1) Confirm that the instrument is at the main menu if it has a keypad/display. If there is an error upon startup, press “stop”, record the error code and press “main menu”. If there is no keypad and the instrument errors upon startup, press the in/out button to stop the chirping before attempting to establish communication.
2) Confirm that the cable is properly attached to the port selected in the software. You can use Microsoft Windows Device Manager to verify the COM port (e.g. COM 1).
3) Confirm that the serial cable and/or USB-serial adapter was obtained from BioTek. Serial cables and USB adapters are not all identical. Contact BioTek customer care to purchase a factory tested cable. After installing a known good cable, reboot the PC, then the reader, then launch the software. Test communication.
4) Confirm that Baud rate is set to 9600 in both the software and the reader. For readers with keypads, confirm the Baud rate under the utilities-setup menu (see Operator’s manual). Reboot the PC, then the reader, then launch the software. Test communication.
5) If steps 1-4 do not resolve the problem, uninstall the suspected COM port from device manager. Reboot the computer to reinstall the COM port.
6) If the reader fails to communicate, and steps 1-5 do not resolve the problem, test the reader on a different PC to confirm which device is at fault. Please contact BioTek if the reader is diagnosed to be faulty.
Your Universal Test Plate comes with a tri-fold paper listing absorbance values at different wavelengths for your plate. (example below) These will be referred to as the "target" values.
For Accuracy measurements, the measurement on your instrument has to lie within specified limits of your target value for that wavelength as follows:
(Target – error) < measurement < (Target + error)
Where error = 1% + 0.01 + instrument specification
The instrument specification is found in the "Specifications" section of your Operator's Manual. It depends on factors such as the read type and plate type and OD of the measurement.
Example: For a Synergy HT, the accuracy specification according to the Operator's Manual in the range of 0.000-2.000 OD for a normal-mode read of a 96-well plate is + 1% + 0.010 OD The total error in this case will be + 2% + 0.020 (the sum of the errors of the instrument and the plate). The target value (from the trifold plate insert) is 0.149 for this example, so the error would be (0.149* 2%) + 0.020 = 0.023.
The passing range would be 0.149 + 0.023, or 0.126 < measurement < 0.172
For Repeatability measurements, you do not use the target value listed on the sheet that came with the plate. For these calculations, the reference is the first reading you make compared to a subsequent reading based on the specifications of your instrument.
Example: For a Synergy HT, the repeatability specification according to the Operator's Manual for a normal-mode read of a 96-well plate in the range of 2.00-3.00 OD is + 3% + 0.005.
For a first read of 2.676, the error would be (2.676 * 3%) + 0.005 = 0.085. The second reading must lie in the range of 2.676 + 0.085, or 2.591 < second measurement < 2.761
There are several options you should consider when having your plates recertified:
* Standard recertification includes the following wavelengths and has a 10-business day turnaround time: 405, 450, 490, 550, 620, 630, 690, and 750 nm.
* Additional wavelengths - for customers working at wavelengths not listed above.
* Precertification data or "as received" data – required by some regulatory bodies.
* Expedited service (2-business day turnaround).
Please contact our service department to receive an RMA and pricing for your test plate.
For biomagnetic separation washing applications such Luminex xMAP bead based assays, several interchangeable, high strength magnet designs are available for the ELx50, ELx405 and EL406 Washers. It is recommended customers work with their BioTek Representative to evaluate the different designs for optimal assay performance.
96- and 384-well Flat Magnets
* Flat-bottom well - beads immobilized in flat band across well bottom
* Round-bottom well - beads pulled to button at well bottom
Note: ~30 µL residual in 96-well plates recommended for best bead recovery.
96- and 384-well Ring Magnets
* Flat-, round-, v-bottom well - beads immobilized in 4-zone ring at well bottom
Note: Enables full well evacuation (residuals <5 µL in 96-well plates). Horizontal dispense offset recommended to properly resuspend beads during dispense cycles.
You can import a text file of “matrix” data (formatted like an 8 x 12 plate, for example) into Gen5 by using the following directions:
1.Before importing a text file, create a protocol and set up an experiment that specifies the exact number of reads that will be imported.
2. Ensure that the delimiters for the text file are the same as those in the Gen5 "System:Preferences:Read From File" settings. The text file format for a semicolon delimited endpoint measurement should look similar to this:
3. Start a new Gen5 Experiment using the protocol designed in Step 1, and click on “Read plate,” choose “Read from File” and browse to find the stored *.txt file to import.
Refer to Gen5’s Help System for information about other importable file formats. Search on “Import Data."