Warranty This warranty may vary outside of the continental United States. Contact your local BioRad office for the exact terms of your warranty. Bio-Rad Laboratories warrants to the customer that the Model 550 Microplate Reader (catalog number 170-6750) will be free from defects in material and workmanship, and will meet all performance specifications for the period of 1 year from the date of shipment. This warranty covers all parts and labor.
Table of Contents Section 1 Introduction ..................................................................................................1 1.1 Accessories to the Model 550 Microplate Reader.....................................................1 Section 2 Product Description .....................................................................................2 2.1 2.2 2.3 Contents of Shipping Carton......................................................................................
Section 1 Introduction The Model 550 Microplate Reader is an eight-channel, vertical pathlength photometer that measures the absorbance of the contents of the wells of 96-well microtitration plates. It can perform single or dual wavelength measurements, and can report absorbance values to three decimal places. The Model 550 reader can be programmed by entering commands through the membrane keypad to define assay blanks, select assay filter(s), set analysis parameters, and select report types.
Section 2 Product Description 2.1 Contents of Shipping Carton The shipping carton contains the following items: 1. Model 550 Microplate Reader, with 415 nm interference filter and 490 nm interference filter installed on the filter wheel 2. Power cord 3. One roll of thermal printer paper 4. Spare fuses 5. Dust cover 6. Instruction manual 7. Warranty card The two additional filters ordered with the instrument are packaged separately and are easily installed by the customer. (See Section 5.1).
13 7 8 9 11 10 12 Fig. 2. Rear View 7. 8. 9. 10. RS-232 serial interface ON/OFF switch Power cord receptacle Fuses 11. Serial Number Label 12. Cooling Fan 13. Cooling Vents 2.3 Membrane Keypad 8 7 1 6 2 5 3 4 Fig. 3. Keypad 1. START/STOP Initiates plate reading using current analysis mode. Stops plate reading. 2. PAPER FEED Advances paper strip in thermal printer. 3. UP ARROW Decreases by one the number on the cursor position or moves a cursor to the preceding mode. 4.
Section 3 Instrument Set-up 3.1 Initial Start-up 1. Place the instrument on a clean, sturdy table or bench. It is important to keep the instrument in a clean, relatively dust free environment to ensure maximum performance. 2. Connect the power cord to the back of the instrument. Before connecting the instrument to the main electrical supply, check that the AC voltage is appropriate for the instrument. 3. Turn on the power switch on the rear panel.
In general, the on-board software is operated by: * Pressing MODE and the ARROW keys until the desired mode is displayed. * Pressing the ARROW keys to choose the parameter to change. * Pressing SELECT to toggle amoung choices. * Pressing ENTER to seal the user’s selection. Calculations The Microplate Reader uses Beer’s Law to calculate the absorbance value of each well.
The first time the instrument is turned on, or after a battery failure, the following default information will be held in the memory. 1. 96 absorbance values of 0.000 2. Dual wavelength reading 3. Measurement filter position is #1 and Reference filter position is #2 4. Plate mixing is inactive (No) 5. Raw data report is selected, all other types are deselected 6. No blank wells are assigned 7. Upper limit is 3.500 and the Lower limit is 0.000 8. The constant type of cutoff report is selected 9.
Notes: 1. If an out-of-range absorbance is measured in one of the blank wells, then the report will show: Blank mean *.*** Std. Dev. *.*** If the out-of-range value is below -3.500, then the asterisks will be preceded by a negative sign (-). C. 2. If the number of blanks is zero, then the report will show: Blank mean 0.000 Std. Dev. 0.000 3. If the number of blanks is one, the report will show: Blank mean Raw absorbance value of the one blank well Std. Dev. 0.
Notes: 1. If one of the positive or negative control wells has an out-of-range absorbance value, the values reported for positive and negative mean and standard deviation will be asterisks. If the out-ot-range value is below -3.500, then the asterisks will be preceded by a minus sign (-). For example, the report may show Pos. Mean *.*** Pos. Dev. *.*** Neg. Mean *.*** Neg. Dev. *.*** Cutoff 1.234 2. If the calculated cutoff value is greater than 3.500 or less than -3.500,then the report will show Cutoff *.
Stored Plate Format No. 1 No. of Standards 0–7 Standard Replicates 1 Sample Replicates 1 Max. No. Samples 88 No. 2 0–7 1 2 44 No. 3 0–3 2 1 88 No. 4 0–3 2 2 44 No. 5 0–7 2 1 80 No. 6 0–7 2 2 40 Format No. 1: Format No.
Format No. 3: Format No.
Format No. 5: Format No. 6: Notes: 1. Blank wells that have been defined under the SET BLANKS mode are replaced by those of the templates. In order to set blanks in the SET BLANKS mode, the number of standards and the number of samples must be set to zero in the SET CONCENTRATION mode (see Section 4.3). 2. If the standards are set to zero, then an error message will be printed on the fourth line of the Concentration report, ERROR: STDs=0. 3.
4. Under three different sets of conditions, an error may be made in calculation of the calibration curve and an error message will be printed on the Concentration report, ERROR: Calibration Curve. Despite the error message, the software does make a calibration curve and uses it to report concentrations. In all three cases, it probably indicates an error was made in preparation of one or more of the standards. a. If the absorbance of any standard is measured as a negative value. b.
4.2 Quick Guide—Reading a plate: 1. Open the rear compartment and verify that the correct interference filters are properly installed in the filter wheel. See Section 5.1 for instructions on changing the interference filters. Close the rear cover. 2. Turn on the instrument. Allow one minute for the self diagnosis. Wait three minutes before reading a plate. While the instrument is warming up: 3. Enter the SET ANALYSIS mode (Section 4.3) to: a. Choose single or dual wavelenth measurement b.
4.3 Detailed Operation Set Analysis Mode In this series of screens the user chooses single or dual wavelength measurement, assigns the measurement and reference (for dual wavelength measurement) filters, and selects or deselects mixing. 1. Enter the SET ANALYSIS mode by pressing MODE until the display reads >PRINT REPORT MODE 2. Press either ARROW until the display reads >SET ANALYSIS MODE 3. Press ENTER. The display will read Set Analysis ■ D/S, ■ Filt and ■ Mix 4. Dual/Single Wavelength Setting: a.
5. Filter selection: a. Use either ARROW to move the cursor under the check box next to Filt. b. Press SELECT to darken the check box next to Filt. c. Press ENTER. The display changes to read Set Filter: Mes.=#x Ref.=#y where x and y refer to filters 1, 2, 3, or 4. d. The cursor will be under the Measurement filter. Use either ARROW to select the filter to be used for measurement. e. Press NEXT to move the cursor to the Reference filter position.
Set Report Types Mode Choose the reports to be generated and automatically printed from within this mode. 1. Press MODE until the display reads >PRINT REPORT MODE 2. Press either ARROW until the display reads >SET REPORT TYPES MODE 3. Press ENTER. The display changes to read ■ Raw ■ Abs ■ Limit ■ Matr ■ Cut ■ Conc 4. To select a report type, use either ARROW to move the cursor under the check box associated with the report type, then press SELECT. The check box will darken.
3. Press ENTER. The display changes to read ■ Clear Blanks ■ Col ■ Row ■ Well Note: If the display instead changes to Err: Conc. Format Press any key. it is because the parameters have been set for a Concentration report and the locations of the blanks cannot be changed. In order to set blanks, go into the Set Concentration mode and set both the number of samples and number of standards to zero (see Section 4.3 Set Concentration). 4. All previously defined blanks can be cleared simultaneously. a.
d. If a column has previously been defined as blank, its check box will be darkened. If a column is not yet defined as blank, then its check box is empty. e. To change the definition of a column (either to ‘blank’ or to ‘no longer blank’), use either ARROW to position the cursor beneath the check box associated with the desired column. f. Press SELECT to darken empty check boxes or to empty darkened ones. g. Press ENTER to complete the selection of the blank columns and to return to the previous screen.
e. When the desired row is chosen, press NEXT to move the cursor to the column position. f. Again, use either ARROW to step up or down between 1 and 12 until the desired column number is displayed. g. The check box on the bottom line of the display will be darkened if the chosen well is presently defined as blank. If the well is not presently defined as blank, then the check box is empty. To change the status of a well, press SELECT.
4. The cursor begins under the first digit of the Upper limit. Use either ARROW to change the first digit. 5. When the first digit is set, press NEXT to move the cursor to the first decimal position. Set the first decimal digit with the ARROW. Continue moving to the right with NEXT until all four digits of the Upper limit are set. 6. When the last digit of the Upper limit is set, press SELECT to move the cursor to the first digit of the Lower limit. 7.
5. In this screen, choose the type of cutoff report desired. a. Use either ARROW to move the cursor under the appropriate check box. b. Press SELECT to darken the check box. c. Press ENTER to seal the selection. The display will revert to the previous screen ■ C/F ■ Set Const ■ Set Formula 6. If the choice in step 5 was Formula, skip to step 9. If the choice in step 5 was Const: a. Move the cursor under the check box next to Set Const. b. Press SELECT to darken the check box. c. Press ENTER.
10. If it is not necessary to change the number of controls, skip to Step 12. To change the number of positive and negative controls: a. Use either ARROW to move the cursor to the check box next to STDs. b. Press SELECT to darken the check box. c. Press ENTER. The display will become Set Formula: ■ STDs = x (0~8) where x is the current number of controls 11. Use either ARROW to change the number of controls to any number between 0 and 8, inclusive. Press ENTER to seal the selection.
14. Negative control locations are defined just as those for positive controls. a. Use either ARROW to move the cursor under the check box next to Neg. b. Press SELECT to darken the check box. c Press ENTER. The display will change to Set Neg.: #1/y well A-1 again where y is the number of controls. 15. Follow the steps as in 13 to define the positions of the negative controls. When finished, press ENTER and the display reverts to Formula: #x ■ STDs ■ Pos ■ Neg 16.
d. First set the number of standards by using either ARROW to move the cursor under the check box next to STDs. e. Press SELECT to darken the check box. f. Press ENTER. The display will change to Set STD: STDs = x (0~7) where x is the currently selected number of standards. g. The cursor will be under the x position. Use either ARROW to change the number to the desired number of standards (0 - 7) and then press ENTER. The screen changes back to ■ STDs ■ Std Repl ■ Std Conc h.
q. Press NEXT to move the cursor to the second digit. Use either ARROW to set the second digit. r. Repeat step q until all four digits are set correctly. s. Press NEXT to return the cursor to the # position. Use either ARROW to change the ‘1’ to ‘2’. t. Continue using NEXT and ARROW to set the concentrations of each standard.
j. To set the number of replicates for each sample, use either ARROW to move the cursor under the check box next to Sample Repl. k. Press SELECT to darken the check box. l. Press ENTER and the screen will change to Set STD Repl: REPL = y (1~2) where y is the currently set number of sample replicates. m. Use either ARROW to set y to the desired number of replicates and then press ENTER twice to return the screen to ■ E STD ■ E Sample ■ Print Format 6. Printing the Concentration format.
c. Press ENTER and the screen will change ■ Print Reports ■ System Config. d. Use either ARROW to move the cursor under the check box next to Print Reports. e. Press SELECT to darken the check box. f. Press ENTER and the printer will begin printing. When printing is completed the screen returns to >PRINT REPORT MODE 2. Printing the system configuration. a. Press MODE until the display shows >PRINT REPORT MODE b. Press ENTER and the display changes to ■ Print Reports ■ System Config. c.
Section 5 Instrument Service by the User Caution: Electrical shock hazard! Always disconnect the instrument from the AC power source before opening the service compartment. Caution: The lamp may be very hot! Use caution, and allow the lamp to cool before attempting service. 5.1 Installing Interference Filters 1. With the power off, open the rear cover of the instrument by sliding the release latch on the top rear of the unit toward the rear of the unit. 2.
Section 6 Troubleshooting and Error Messages The Model 550 Microplate Reader constantly monitors several vital functions and will display appropriate messages on the LCD when an error condition is encountered. 1. LCD appears blank on power up. a. Power switch is not turned on. Turn on the power switch on the rear panel. b.Unit is not plugged into AC outlet or power cord is not attached to the instrument. Check power cord connections at outlet and on the rear panel of the instrument. c.
b.The best measurement wavelength filter for the substrate in use has not been installed in the filter wheel. Check references for the substrate you are using, or determine the best measurement wavelength by analyzing the substrate product on a scanning spectrophotometer. Custom filters are available from 400 to 700 nm. c. The filters have been switched in the wheel. Check that the filters are installed in the correct position. 7.
Section 7 Specifications 7.
7.2 RS-232 Interface Specifications The following specification describes the syntax and language required for computer control of the reader. In addition, the Model 550 reader will perform passive unidirectional data transmission. The instrument will automatically transmit the data in ASCII code after each reading if it is properly connected to a host computer. See the “RTPLATE” command for an example of ASCII output. Note: This automatic output will be of Measurement O.D. minus Reference O.D.
7.3 Command Language for the Model 550 Microplate Reader SYNTAX:[ ] device name: “EIA. READER” -Model 550 Microplate READER 1. : “ID”-request device ID. 550 READER id; “550”; no arguments : “ERE” : “0550”: Model 550 Microplate READER ID code] 2.
8. : “RPLATE” - read plate; arguments , , : mix - Mixing time before reading (in seconds) ; ASCII encoded decimal (0-9) wp1 - Measurement filter position; ASCII encoded decimal (1-4) wp2 - Reference filter position; ASCII encoded decimal (1-4) Optional; only present for Dual wavelength reading : “ERE” 9. : “RTPLATE” - retransmit the last plate read; no arguments : “ERE”
Error Codes “0000” - No error “8071” - Invalid Command “8072” - Parameter Out of Range “8073” - Device Not In Remote Mode “8074” - Device Busy “8075” - Not assigined “8076” - Not assigined “8077” - Light Bulb Burned Out “8078” - Hardware Error “8079” - Memory Error “8080” - Not assigined “8081” - Not assigined Note: Until the Model 550 Reader receives the “AQ” command, the reader will be unable to accept any other command and will only respond with the error code of “8073”- Device Not In Remote Mode.
Bio-Rad Laboratories Life Science Group 4000067 BulletinRev 0000D US/EG Rev A Web site www.bio-rad.com Bio-Rad Laboratories Main Office 2000 Alfred Nobel Drive, Hercules, CA 94547, Ph. (510) 741-1000, Fx. (510) 741-5800 Also in: Australia Ph. 02 9914 2800, Fx. 02 9914 2889 Austria Ph. (01) 877 89 01, Fx. (01) 876 56 29 Belgium Ph. 09-385 55 11, Fx. 09-385 65 54 Brazil Ph. 55 21 507 6191 Canada Ph. (905) 712-2771, Fx. (905) 712-2990 China Ph. 86-10-8201-1366/68, Fx. 86-10-8201-1367 Denmark Ph.
