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It is important to note that when measuring absorbance in a microplate well, the pathlength is 
determined by the volume. In a microplate assay, it is very important that all wells have the same 
volume and that all solutions are dispensed with an accurate liquid handler like an 8-channel pipette. 

Common applications of absorbance-based microplate readers are ELISA, total protein (like Bradford 
or BCA) and cell proliferation assays. 

 

2

 

Instrument Use 

The Biochrom EZ Read 800 Plus Microplate Reader is used for microplate-based applications 
requiring endpoint or kinetic absorbance measurements from 400-750 nm in optically clear 96-well 
plates with a standard SBS/ANSI footprint. 

Only trained laboratory personnel should operate the Biochrom EZ Read 800 Plus Microplate 
Reader. The Biochrom EZ Read 800 Plus Microplate Reader is intended for general laboratory and 
research use only.  

 

 

 

 

Содержание 800 Plus

Страница 1: ...1 EZ Read 800 Plus Microplate Reader User Manual Biochrom Ltd 22 Cambridge Science Park Cambridge UK CB4 0FJ Tel 44 1223 423723 ...

Страница 2: ... 3 2 SOFTWARE INSTALLATION 6 3 3 QUICK MEASUREMENT 7 3 4 KINETIC MEASUREMENT 12 3 5 MULTI WAVELENGTH MEASUREMENT 17 3 6 CALIBRATING THE INSTRUMENT 20 3 7 USING THE USER INTERFACE ON THE EZ READ 800 PLUS 20 4 TECHNICAL INFORMATION 39 5 GENERAL FEATURES 39 5 1 MEASUREMENT MODES 39 5 2 INTEGRATED QUALITY CONTROL 39 5 3 SCOPE OF SUPPLY 40 5 4 LIABILITY 40 6 MAINTENANCE 40 6 1 APPROVED PARTS 40 6 2 CLE...

Страница 3: ...IONS 41 7 2 RETURNS TERMS AND CONDITIONS 42 7 3 RETURNS PROCEDURE 43 8 ORDERING INFORMATION AND ACCESSORIES 45 9 CONTACT INFORMATION 45 9 1 TABLE 2 CONTACT INFORMATION 46 10 APPENDIX 46 10 1 HELPFUL HINTS FOR SUCCESSFUL MICROPLATE ASSAYS 46 10 2 TROUBLESHOOTING AND FREQUENTLY ASKED QUESTIONS 46 11 DECLARATION OF CONFORMITY 49 ...

Страница 4: ...ction What is a microplate reader A microplate reader is a used to measure the absorbance of liquid samples in a 96 welll plate What do microplate readers measure Microplate readers measure the amount of specific wavelengths of light that is absorbed by molecules within a solution Why is this useful and how do microplate readers measure sample absorbance The amount of light absorbed by molecules w...

Страница 5: ...croplate readers are ELISA total protein like Bradford or BCA and cell proliferation assays 2 Instrument Use The Biochrom EZ Read 800 Plus Microplate Reader is used for microplate based applications requiring endpoint or kinetic absorbance measurements from 400 750 nm in optically clear 96 well plates with a standard SBS ANSI footprint Only trained laboratory personnel should operate the Biochrom ...

Страница 6: ...ia the supplied USB cable 2 To connect the instrument to Galapagos software Insert the CD or USB supplied with the instrument into a PC Galapagos software includes pre requisite files dot NET Framework 4 Windows Installer and Microsoft SQL Express 2008 which allow installation to occur Insert the CD or USB and run the setup file A pop up menu will appear and guide you through automatic installatio...

Страница 7: ... measurement at one wavelength c Dual wavelength refers to the measurement of a sample at two wavelengths The first wavelength is where the sample of interest absorbs The second wavelength is used as a reference and will be subtracted from the first wavelength The absorbance measurement is then subtracted from a measurement at a different wavelength Multiple wavelength refers to a series of absorb...

Страница 8: ...elect to shake the microplate Users can select shaking to be a single event or in timed cycles 4 Select the amplitude degree of linear shaking 5 Select the speed for linear shaking 6 Users can select the duration of shaking ...

Страница 9: ...lick Start to begin acquisition 8 Raw absorbance values for dual wavelengths measurements are displayed in the wells Details of corresponding wavelengths can be displayed when clicking on the individual well ...

Страница 10: ... values are displayed The bold value refers to the absorbance measurement after wavelength subtraction of the absorbance at the measurement wavelength from the absorbance at the reference wavelength The second value is the absorbance at the measurement wavelength The third value is the absorbance at the reference wavelength ...

Страница 11: ...xternal database system such as LIMS Users can save files and templates onto the database File save to own personal file or Save Method Template saves the method protocol for future use 11 Results can also be exported as Excel Extended Metafile results exported as a HTML file PDF Rich Text File Text File Word File and XPS File ...

Страница 12: ...s measurements click on the timer icon for timed measurements This is indicated by Timing On Users can then input the required kinetic parameters for a measurement cycle The measurement cycle refers to the point of absorbance measurement including any timed delay and or interval until the next reading Note the use of a delay is optional tick or un tick whereas the interval and the number of cycles...

Страница 13: ...le Dual or Multiple wavelength modes Note measuring at more than one wavelength will affect the interval that can be used The interval time must take into account the measurement time at a single wavelength e g this is 5 seconds at a single wavelength for the EZ Read 800 Plus Note Method details of the kinetic experiment are listed on the left hand panel of the screen ...

Страница 14: ...elect to shake the microplate Users can select shaking to be a single event or in timed cycles 7 Select the amplitude degree of linear shaking 8 Select the speed for linear shaking 9 Users can select the duration of shaking ...

Страница 15: ...15 The lines shown in each well refer to the measurement at one wavelength over time Different wavelengths used for absorbance measurements are assigned a specific colour and identified by a legend ...

Страница 16: ... external database system such as LIMS Users can save files and templates onto the database File save to own personal file or Save Method Template saves the method protocol for future use 11 Results can be exported as Excel Extended Metafile results exported as a HTML file PDF Rich Text File Text File Word File and XPS File ...

Страница 17: ...e sample s composition or purity 1 To perform a multi wavelength measurements click Multiple Wavelength from the drop down menu 2 The menu is adjusted to allow users to select up to 4 wavelengths for measurement Each wavelength must be different For the EZ Read 800 Plus ELISA users can input wavelengths of 405 450 492 and 620nm For the EZ Read Research model users can choose four wavelengths from ...

Страница 18: ...in each well with raw absorbance displayed on the right hand panel as a graph of absorbance vs wavelength 4 Alternatively users can click on an individual well to view the scan in more detail and raw results Both tabulated and graphical chart data are displayed ...

Страница 19: ...ase system such as LIMS Users can save files and templates onto the database File save to own personal file or Save Method Template saves the method protocol for future use 6 Results can also be may be exported as file types such as Excel Extended Metafile results exported as a HTML file PDF Rich Text File Text File Word File and XPS File ...

Страница 20: ...al liquid crystal display provides the user with set up parameters and experimental results The keypad is used for navigation and has a spill proof membrane Navigation using the on board menu is very simple 1 Press the corresponding number on the keypad to enter the user mode choices or numeric data 2 Press keypad directly below the corresponding option on the display F1 F2 F3 and F4 to select the...

Страница 21: ...method to use 3 Define Method This option enables a new method to be defined Method Definition either from the beginning or by modification of a previously stored or default method 4 Recall Plate Data This option recalls previously stored measurement data 5 Setup This option enables the basic plate reader parameters such as printer driver and filter set up calibration to be set up 3 7 2 Method Def...

Страница 22: ...e entries 4 Kinetics Press F4 if kinetics is required Enter the interval time time between readings and the number of measurements The total process time is calculated and indicated on the screen The minimum interval time is 10 seconds and the maximum number of measurements is 100 NOTE Up to 100 plate data results can be stored in the instruments memory If during the kinetic measurement the maximu...

Страница 23: ...ant them to be They are not tied For example LP and HP can be used to substitute for terms T1 and T2 defined in a specific test kit The terms can be used in calculations defined later in the method definition BK Blank The absorbance of the blank value s is automatically subtracted from the absorbance of the other wells NC Negative control Used for qualitative screening tests PC Positive control Us...

Страница 24: ...epeat pressing of F4 as appropriate By alternating between F1 and F4 it is possible to build up the plate as required You can come back to edit the layout if required Use the arrow keys to move around the plate layout Above the plate illustration the current position and well type is shown 4 Standards When ST standards is selected ST1 option appears as F2 After placing these on the layout press F2...

Страница 25: ...ples Place the first sample SM1 and replicates using F4 Press F1 to get SM2 Use F4 to place the second sample and repeat as necessary Replicates of samples can be placed in any free location it is not necessary that the wells are adjacent 3 Fill all To reproduce the first sample pattern for all samples Sample 1 has to be defined The Function Fill All F2 will fill the plate accordingly as set in Op...

Страница 26: ... Quick key F1 offers a quick way of entering the concentrations 3 Units can be defined using the alphanumeric routine 4 Select the analysis model for the standard curve The options are Linear regression Parabolic Cubic Point to Point linear interpolation Spline and 4 parameter 5 Select the y and x axis format required for presentation of the results linear linear log linear linear log log log ...

Страница 27: ...on 1 Five kinetic calculation options are available 1 Onset time absolute This option calculates the time required to reach the absorbance value specified in the input field for each well A linear regression is used to interpolate the time between the kinetic measurement points 2 Onset time delta The option calculates the time required for the Absorbance difference between that of the first measur...

Страница 28: ...nce change normalized to a minute for the three readings at the maximum slope 5 Delta OD The absorbance change normalized to a minute over the whole measurement time is calculated NOTE If none of these kinetic options is chosen the raw absorbance data for each well at the specified time intervals can be printed out 3 7 2 6 Method Definition Calculations 1 If kinetics is not selected the Calculatio...

Страница 29: ...cify the use of a factor or factors to normalize results to what they should be 2 These factors can be used in any further calculation step e g in the transformation formulas 3 Press enter to move to the next method definition screen 3 7 2 8 Method Definition Elimination This option allows users to define conditions for the automatic elimination of individual controls which do not meet the set cri...

Страница 30: ...e positive control is lower than 85 of the mean value then eliminate the respective single positive control 5 When the cursor is in the entry field the function keys are active and users can select different options There are three groups of elements that can be selected by pressing the F1 Alt key Controls single and average and the factors Operates etc All available mathematical and logical funct...

Страница 31: ... to select the desired element and F4 to place it NOTE If a Blank BK is defined it will be automatically subtracted from the absorbencies of all other wells 3 Example An index in per cent based on the absorbance of the CO control should be calculated for each well Absorbance of the CO control 100 The formula should be entered as follows X 100 avg CO 4 Press enter to move to the next method definit...

Страница 32: ...he threshold s proceed as follows Select the data source first The options are Val absorbance or kinetic data Trf transformed data Conc concentration Next enter the label for the upper limit in the top left edit field This can be a text like Positive or a symbol like Now enter the threshold condition for the upper limit This can be a value or an equation After the condition for the upper limit has...

Страница 33: ...is is indicated on the printout Validation criteria can be defined for the measured absorbance values or kinetic values Val transformed values Trf as defined in Transformation or concentration values Conc depending on the test protocol 1 To define a validation condition chose the data source first Then enter the first condition At the bottom of the screen you can see various options like avg PC Fc...

Страница 34: ...ion Printout Use this option to select the content of the printout You can select one several or all options Only options relevant to the assay that has been defined selected are displayed The image below shows all options that are possible if kinetics has been selected the option for absorbances is replaced by Kinetic values 1 If replicates have been used the average value can be printed out 2 Pr...

Страница 35: ... immediately after defining a method press 1 or by recalling an existing method press 2 select using and F4 The plate carrier will move out now allowing loading the microplate 1 Each plate requires a unique ID This ID is used to identify the plate on the printout and the measured data for later recall 2 In case an ID already used is entered the screen will show a warning asking if the previous mea...

Страница 36: ...eading and subsequent printing Up to 100 plate data results can be stored in memory After this is full the new results overwrite the previous results To avoid this results can be downloaded to PC for archiving 3 7 4 Recalling a plate Previously run assays are stored using the unique plate ID The Recall menu can be called from the main menu by pressing key 3 and the ID 1 The screen below shows a li...

Страница 37: ... values obtained for the plate No calculations are carried out NOTE It is the sole responsibility of the user to make sure that the chosen method gives useful results The program cannot determine if the selected method makes sense with the existing data 4 When the measured OD values should be printed press the key F3 Raw No calculations will be performed with this option 5 To show the measured OD ...

Страница 38: ... compatible type Use the up down arrow keys to highlight the printer required and press enter to confirm your selection Depending on the printer type some options like paper size may be shown Use the function keys to select the required option and press the OK key to confirm the selection 3 7 5 2 Setup Filters The Filter submenu serves to review the installed filters inform the system about new or...

Страница 39: ... display using the F1 and F2 keys 3 7 5 5 Setup Language Use to select the language that is used on the displays and printouts 4 Technical Information Please see Biochrom s website www biochrom co uk for complete technical specifications 5 General Features 5 1 Measurement modes Single dual and multi wavelength and kinetics measurements with absorbance data output in OD on display 5 2 Integrated qu...

Страница 40: ...e affected Biochrom is not liable for damages or costs caused by unauthorized alterations repairs or modifications of the equipment Maintenance 6 1 Approved Parts Except for the parts shown in the following list only parts supplied by Biochrom or an authorized Biochrom Distributor may be installed in or used with the EZ Read 800 Plus 6 2 Cleaning and Disinfection All parts of the reader that come ...

Страница 41: ...and let it soak for 30 minutes 4 When a bleach solution has been used carefully wipe off the entire reader with lint free tissues that have been moistened in water 5 Dry the reader by wiping it off with lint free tissues 6 4 Additional Maintenance Tips Never spray liquids directly into the interior of the reader Do not use force when cleaning the reader Wipe up spills immediately Avoid buildup of ...

Страница 42: ...ltered Biochrom EZ Read 800 Plus Microplate Reader has not been installed in accordance with supplied instructions Biochrom EZ Read 800 Plus Microplate Reader has been damaged during return transport due to wrong packing e g not in original packing material Biochrom EZ Read 800 Plus Microplate Reader was damaged due to improper operation and outside of the use described in this manual If an instru...

Страница 43: ...ontamination certificate from and include this certificate with your package Please put a copy on the outside and inside of your package Please Note Instruments which are returned without a decontamination certificate will not be processed This would result in significant delays and additional administration charges for your repair IMPORTANT For returns outside the EU please complete your commerci...

Страница 44: ... value Customs declaration Temporary Import etc Have read and understood the returns terms and conditions http www biochrom co uk returns_terms html For all returns enquires please contact returns biochrom co uk or 44 0 1223 427861 ...

Страница 45: ...for a list of filters that are available for purchase 9 Contact Information Table 2 Contact Information Department Email contact Telephone contact Sales enquiries biochrom co uk 44 0 1223 423723 Technical Support support biochrom co uk 44 0 1223 427890 Orders orders biochrom co uk 44 0 1223 427861 44 0 1223 420 164 Fax Returns returns biochrom co uk 44 0 1223 427861 Human Resources hr biochrom co ...

Страница 46: ...which your molecule of interest does not absorb Many assays recommend a reference measurement at 620 nm When using plate frames for stripwells or single wells breakable strips check that all strips and wells are pushed down completely and are level with the frame Keep the bottom of the plate dry If liquid has contaminated the well bottoms or if condensation is present dry the surface with a lint f...

Страница 47: ...located to certain controls such as positive or negative controls Users can input details of the controls such as the name and type of control Users can use threshold calculations to subtract readings from other wells 4 Which Microsoft Windows operating systems are Galapagos compatible with Galapagos is compatible with Windows XP with SP3 and higher versions 5 Why do I get different readings from ...

Страница 48: ...ly used reference wavelength in many absorbance applications in the visible range Check the product data sheet for recommended reference wavelength for your specific assay 6 How can I connect the instrument to Galapagos Refer to section 3 2 ...

Страница 49: ...ared are as follows EN61010 1 2010 Safety requirements for electrical equipment for measurement control and laboratory use General requirements EN61010 2 101 2002 Safety requirements for electrical equipment for measurement control and laboratory use Particular requirements for in vitro diagnostic IVD medical equipment EN61326 1 2006 Electrical equipment for measurement control and laboratory use ...

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