Bio-rad Microplate Manager Software Uživatelský manuál

Microplate SystemsMicroplate Manager® 6 SoftwareInstruction Manual | Version 6.1

Introduction 2 Microplate Manager 6 can perform four types of microplate readings. • Endpoint Reads are used to acquire a single absorbance reading

Appendix 92 Percent Max Binding Assay Tutorial 1 Defining the Template 2 Setting up the Equations 3 Selecting Curve Fit and Customizing Curve

Appendix 93 • Blanks are defined as #. The average of these # wells will be subtracted from all the wells. • NSB are non-specific binding wells. •

Appendix 94 3. Click Template View icon in the Template Toolbar 4. Select Equation from popup 5. To apply this equation to the data, right click

Appendix 95 3. Select 4-Parameters from list 4. Click Customize Plot icon in Curve Fit Plot Toolbar 5. In the Curve Fit Plot Editor window under

Appendix 96 6. In the Curve Fit Plot Editor window under X-axis tab, enter Concentration for the axis label. Set the Range to Manual with the minimum

Appendix 97 7. In the Curve Fit Plot Editor window under Y-axis tab, enter % B / Bo for the axis label. Set the range to Manual with the minimum 40.0

Appendix 98 8. The customized curve fit plot shows the curve fit parameters below the curve.

Appendix 99 4 Entering Experimental Info 1. Click on the Info icon on the Main Toolbar 2. Enter Title of assay and any experimental Comments. 3.

Appendix 100 5 Setting up the Report 1. Click Report icon in Main Toolbar 2. Click Select Report Contents icon in the Report Toolbar. 3. Move the

Appendix 101 5. Select Columns to be included for Standard and Sample Tables in Report window. Move the items up or down to select which items to inc

Introduction 3 File Menu After creating a New Experiment or opening an existing data file, the File Menu will list the main program functions. The ni

Appendix 102 A section of the Data Analysis Report for Standards and Samples Tables with user-defined name % B/Bo as a column header is shown here.

Appendix 103 8. Click the Print Preview icon in the Report Toolbar 9. Click the Print icon in the Print Preview Toolbar to Print the Report

Life Science Group 1208 Sig 030810011683 Rev B US/EGBio-Rad Laboratories, Inc. Web site www.bio-rad.com USA 800 4BIORAD Austr

Introduction 4 • Export Data to Excel • Read a Plate • Print Report/active window • Print Preview of Report Window Menu After creating a New Exper

Installation and Instrument Setup 5 Installation and Instrument Setup Minimum Requirements Computer • Windows XP, Windows Vista or Mac OS 10.4 or hig

Installation and Instrument Setup 6 For Mac Users: 1. Insert the software CD into the CD-ROM drive. The installer user interface displays. 2. Drag t

Installation and Instrument Setup 7 ¾ If filters are added, removed, or their positions changed, the changes must be entered on the iMark reader keyp

Installation and Instrument Setup 8 xMark Instrument Setup 1. After opening MPM 6 for the first time, choose New Experiment from the File menu. 2. S

Installation and Instrument Setup 9 5. Select the Shaker Only tab to set up mixing parameters (xMark only).

Quick Start 10 Quick Start Setting Up an Experiment 1. Launch software from the desktop shortcut. 2. Click on the New Experiment icon or choose Fi

Quick Start 11 Open a saved protocol or set up a new protocol (by setting up a template, and selecting the instrument, assay info, analysis options an

Quick Start 12 Incubator Controls Under the Instrument Control dialog box, select the Incubator tab (xMark only) 1. Click the Set to On button. 2. E

Quick Start 13 Reading Microplates After all Instrument settings have been completed, press the Start Read button to read the plate. Data analysis res

Data and Protocol Files 14 Data and Protocol Files The following table explains how protocol and data files differ, and describes the file types MPM 6

Data and Protocol Files 15 The Sample Data files are stored in the following default location: C: \ Bio-Rad \ Microplate Manager 6 \ Sample Data The

Data and Protocol Files 16 Data AutoSave When the AutoSave feature is selected in the Instrument Setup Window, the data will be saved automatically af

Data and Protocol Files 17 3. Change file Extension from ".epc " to " .csv " In Microplate Manager 6, 1. Choose New Experimen

Data and Protocol Files 18 Data can also be entered into the program by typing from the keyboard. Exporting Data or Reports Both raw data a

Data and Protocol Files 19 The exported file will be saved as an Excel (.xls) file. Exporting Reports 1. To export data in a report format, including

Setting Up a Protocol 20 Exporting Custom Screening Reports (BSE, TSE, TSE, NSP) (Windows PC only, using Bio-Rad custom Excel templates BSE, TSE, TSE(

Setting up a Protocol 21 5. Select File >Custom Export. 6. Select desired Template (which contains the built-in calculation functions for repor

Microplate Manager 6 i Bio-Rad Technical Support The Bio-Rad Technical Support Department in the United States is open Monday–Friday, 6:00 a.m. to 5:

Setting Up a Protocol 22 Section, the Control Validation Section, the Cutoff Values Section, and the Matrix Data including the Sample info. In the Inf

Setting up a Protocol 23 The matrix of the 96-well absorbance data, including the well identifiers, is shown in the fourth section. Positive control

Setting Up a Protocol 24 Setting up a Protocol Setting up protocols automates your test processes. Parameters for frequently-run assays can be set up

Setting up a Protocol 25 Setting Up A Template The template defines the identifier locations for your plate data. A template must always be present in

Setting Up a Protocol 26 To highlight the entire plate, choose Select All under the Edit menu. To clear the template, press the delete key. To re-siz

Setting up a Protocol 27 Entering Dilutions Dilutions can be entered for any sample well (except for blanks or standards). To enter Dilutions: 1. Cli

Setting Up a Protocol 28 Select up to 6 Custom Labels from a pop-down menu by selecting one of the items in the list, or by entering a new Label ID in

Setting up a Protocol 29 3. In the Curve Fit Plot Editor, select Auto/ Manual Scaling, Number Format, Axes labels, etc. for Curve Fit Plot.

Setting Up a Protocol 30 Setting up the Report Contents 1. Click Report Icon in Main Toolbar. 2. Click the Select Report Contents icon, and then,

Setting up a Protocol 31 5. Click Format Report Editor icon in the Report toolbar to format the report with titles, number format, headers and footer

Introduction ii Table of Contents Table of Contents... ii Introduction ...

Analyzing Data Using a Standard Curve 32 Analyzing Data Using a Standard Curve The standard curve is a plot of the concentration of the standards you

Analyzing Data Using a Standard Curve 33 Standard Curve Analysis Options Linear Fit For a linear fit, the best straight line is fitted to the data poi

Analyzing Data Using a Standard Curve 34 Zero-intercept Linear In the Zero-intercept of analysis a linear fit is used, but the intercept is set to Zer

Analyzing Data Using a Standard Curve 35 Semi-log Fit When Semi-Log is selected in the Curve Fit dialog box, a plot of the response (y axis) vs. the l

Analyzing Data Using a Standard Curve 36 Log/log Fit The Log/Log curve fit option fits the best straight line to the data consisting of the logarithm

Analyzing Data Using a Standard Curve 37 Log/Logit and 4-Parameter Fit Both the Log/Logit and the 4-Parameter curve fit methods are based on the same

Analyzing Data Using a Standard Curve 38 The 4-parameter method is usually preferred over the Log/Logit method, because the results for the 4-paramete

Analyzing Data Using a Standard Curve 39 Point to Point In the Point to Point method of analysis, the data points are simply connected. No fitting of

Analyzing Data Using a Standard Curve 40 A minimum number of standards that pass certain criteria are required for each type of curve fit. For the bes

Analyzing Data Using a Standard Curve 41 To obtain a printed standard table report for the imported standard curve, print the imported standard curve

Microplate Manager 6 iii Exporting Data ... 18 Exporting Reports

Analyzing Data Using a Standard Curve 42 Equations and Calculations Definitions of the statistical parameters calculated for a set of values <y>

Analyzing Data Using a Standard Curve 43 A residual is the difference between an observed value of the response variable and the value predicted by th

Analyzing Data Using a Standard Curve 44 Error of the interpolated concentration (concy) is related to the error of the response sy according to s con

Analyzing Data Using Cutoffs 45 Analyzing Data Using Cutoffs The Cutoff Result analysis options are used for assays including one or two controls (and

Analyzing Data Using Cutoffs 46 The three cutoff display options in the menu are described here. Either two or four cutoffs are used based on the use

Analyzing Data Using Cutoffs 47 Reference can be made to standard or sample IDs that have been defined in the Template setup • Standards: enter ! fo

Analyzing Data Using Cutoffs 48 Data transformed – viewed as a Percent (0 - 100 %) • Low Cutoff: 0 • High Cutoff: 100 Using values from a Standard

Analyzing Data Using Cutoffs 49 The Control wells are defined on the template. The resulting Extended Cutoff Report, is shown here with the high value

Analyzing Data Using Cutoffs 50 2. Click the Select Table Columns icon in the Report Toolbar.. 3. In the Columns Selector window, move Ext. Cutoff V

Analyzing Data Using Cutoffs 51 The information entered in the Info window is automatically listed at the top of the Report. Concentration unit inf

Introduction iv Running Kinetic Assays... 52 Reading a Kinetic Assay...

Running Kinetic Assays 52 Running Kinetic Assays Running assays kinetically has many advantages: • Kinetic assays are more accurate. The errors in en

Running Kinetic Assays 53 3. Enter the interval between the start of two consecutive reads in the Interval column. The interval time is the time nec

Running Kinetic Assays 54 Kinetic Zoom Plots Zoom in on a range of wells or a single well in the Kinetic Plots display to get an enlarged view of the

Running Kinetic Assays 55 To zoom in on a single well or return to the display of the whole plate: 1. Right-click in the desired plot. 2. Select Zoo

Running Kinetic Assays 56 After the read is completed, data can be excluded from calculation by defining a new time range or new OD range. Original d

Producing Reports 57 Running Spectra Assays (xMark only) 1. Select File > Instrument Setup, Spectra Reading Mode. 2. Enter desired wavelength ran

Producing Reports 58 Producing Reports Setting Up Reports To create a Report, click on the Report icon in the main toolbar, or select Report under t

Producing Reports 59 Click the Select Report Contents icon in the Report toolbar. Select Report Contents Select Table Columns Format Report

Producing Reports 60 5. Select sorting option, Ascending or Descending Separate tabs for Standard and Samples. 6. Click the Format Report icon in

Producing Reports 61 With Report selected under the Window menu, choose File > Print to print the report. Printing Reports 1. Choose File >

Microplate Manager 6 v 3 Selecting Curve Fit and Customizing Curve Fit Plot...73 4 Entering Experimental Info ...

Producing Reports 62 Or, click on the Report icon in the Main toolbar. 3. After setting up the report, click on Print Preview icon to preview the ent

Appendix 63 Customizing Reports 1. Click on the Report icon on the main toolbar. 2. Click on the Format Report icon on the report window. In the R

Appendix 64 Appendix References [1] Channing Rodgers, R. P. (1984) "Data Analysis and Quality Control of Assays: A Practical Primer" in Pra

Appendix 65 Quantitative ELISA Assay Tutorial A tutorial showing how to set up an ELISA Assay, including a standard curve and equations to transform

Appendix 66 Setting up the Equations 1. Click Template View icon, select Equation from popup. 2. Click Equation Editor icon. Choose % Max from Equ

Appendix 67 Selecting and Customizing a Curve Fit Plot 1. Click Curve Fit Plot icon in Main Toolbar. 2. Choose Fitting Function icon in Curve Fit Pl

Appendix 68 Dilution Data Assay Tutorial 1. Defining the Template 2. Setting up the Dilutions 3. Selecting Curve Fit and Customizing Curve

Appendix 69 5. Enter as below and click OK 6. Type 0 in well H1 & H2 and press Enter. 7. Type S01 in well A3 and press Enter. 8. Highlight

Appendix 70 11. Below is the finished template o Standards are defined 200, 100, 50, 25, 12.5, 6.25, 3.125, & 0. o S01 to S20 are all of the u

Appendix 71 3. Highlight area well A3-D3. (Click & hold mouse on well A3, drag mouse to well D3, and release.) 4. Click on the Fill Series icon

Appendix 72 9. Click on the Paste icon in the Template Toolbar 10. E3 to H3 are filled 11. Highlight area well A3-H12. (Click & hold mouse on

Appendix 73 3 Selecting Curve Fit and Customizing Curve Fit Plot 1. Click Curve Fit Plot icon on the Main Toolbar 2. Click Fitting Function icon i

Appendix 74 5. In the Curve Fit Plot Editor window under Plot tab, select the Color, Symbol, and Line style for the Data Points and Curve Fit Line.

Appendix 75 6. In the Curve Fit Plot Editor window under X-axis tab, enter Concentration for the axis label. Set the Range to Auto. 7. In the Curve

Appendix 76 8. The customized curve fit plot shows the curve fit parameters below the curve

Appendix 77 4 Entering Experimental Info 1. Click on the Info icon on the Main Toolbar 2. Enter Title of assay and any experimental Comments. 3.

Appendix 78 5 Setting up the Report 1. Click Report icon in Main Toolbar 2. Click Select Report Contents icon in the Report Toolbar 3. Move the i

Appendix 79 5. Select Columns to be included for Standard and Sample Tables in Report window. Move the items up or down to select which items to incl

Appendix 80 A section of the Data Analysis Report for Standards and Samples Tables with Average Concentration is shown here.

Appendix 81 6. Click the Print Preview icon in the Report 7. Click the Print icon in the Print Preview Toolbar to Print the Report

Introduction 1 Introduction This manual assumes that you are familiar with the standard commands and functions associated with the Windows® or Mac® op

Appendix 82 Percent Control Assay Tutorial 1 Defining the Template 2 Setting up the Equations 3 Setting up the Screening Report 4 Entering

Appendix 83 • CNTRL is for the control wells. • U01 to U29 are all of the unknown samples to be analyzed. 2 Setting up the Equations 1. Click Equat

Appendix 84 3. Click Template View icon in the Template Toolbar. 4. Select Equation from popup. 5. To apply this equation to the data, right clic

Appendix 85 3. Define the desired cutoffs in the Cutoffs Report Editor window. Well identifiers used in the Cutoff Report Editor must be first define

Appendix 86 4. Click the Report View icon in the Result Toolbar 5. Select Extended Cutoff Report from popup 6. The resulting Extended Cutoff Repo

Appendix 87 4 Entering Experimental info 1. Click on the Info icon on the Main Toolbar 2. Enter Title of assay and any experimental Comments. 3.

Appendix 88 5 Setting up Report 1. Click Report icon in Main Toolbar 2. Click Select Report Contents icon in the Report Toolbar. 3. Move the item

Appendix 89 5. Select Columns to be included for Sample Table in Report window. Move the items up or down to select which items to include in the Rep

Appendix 90 A section of the Data Analysis Report for Samples with user-defined name % CNTRL as a column header is shown here. 8. Click the Print P

Appendix 91 9. Click the Print icon in the Print Preview Toolbar to Print the Report