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Posted by aliu under MasterPlex ReaderFit

Dose response assays are generally characterized by a nonlinear relationship between the response and the amount of drug/agonist/antagonist given. When plotted, this response will usually exhibit a sigmoidal curve that is best described using the 4 Parameter Logistic (4-PL) or 5 Parameter Logistic (5-PL) nonlinear regression model equations.

Dose Response Curve

Dose Response Curve

As described in our Bioassay Analysis with the Five Parameter Logistic (5-PL) Non-Linear Regression Curve-Fitting Model blog post, the C parameter in both the 4-PL and 5-PL models represent the concentration value for the inflection point of the curve. Depending on whether you have an inhibition or activation assay, the C parameter also represents the half maximal inhibitory concentration (IC50) or the half maximal effective concentration (EC50) respectively.

Anatomy of the 4 or 5 Parameter Logistic Model

Anatomy of the 4 or 5 Parameter Logistic Model

MasterPlex ReaderFit offers full support for the 4 Parameter Logistic (4-PL) and 5 Parameter Logistic (5-PL) model equations making it an ideal tool for automated fitting of dose response curves and calculating the EC50 or IC50 values.

Here is a Quick Guide for this procedure:

  1. To acquire MasterPlex ReaderFit, download and install the free 14-day trial.
  2. If you have not yet imported your data, please refer to the community post on importing data into MasterPlex ReaderFit.
  3. This is a sample dose response data set which contains 3 different sets of data from which we will generate 3 dose response curves (each outlined with a different color). Each curve set contains 8 data points with duplicates that are side-by-side.
    Sample Dose Response Data

    Sample Dose Response Data

  4. Select 1 entire set of data points (including replicates) and click the blue Standard button. Please note that you can hold down the CTRL button and left click non-contiguous wells.

    Marking Data Sets

    Marking Data Sets

  5. The Auto Fill dialog will pop up. This feature is only helpful if you have a serial dilution for your drug concentrations. If not, you will have to enter them in manually.

    Auto Fill Feature for Entering in Serial Dilution Concentrations

    Auto Fill Feature for Entering in Serial Dilution Concentrations

    In this example, Drug ABC was serially diluted from top (20,000 pg/mL) to bottom (0.26 pg/mL) with a dilution factor of 5.

  6. Click Apply & View to preview the values in plate tab to confirm.
  7. Repeat steps 2-4 for any other data sets you may have. In this example, there are 2 other data sets.
    For each cell, you will notice 2 numbers: the response value on the top and the drug dose on the bottom.
  8. Click on the Standard Curve tab.
  9. Make sure the Use Best Fit feature checkbox is unchecked and select the model equation (4-PL or 5-PL) you would like to use for the curve fit and then click Calculate.
    Model Equation Selection and Calculation

    Model Equation Selection and Calculation

  10. MasterPlex ReaderFit will then draw the dose response curve(s) according to the parameters you have chosen.
    Dose Response Curves based on the 4 Parameter Logistic

    Dose Response Curves based on the 4 Parameter Logistic

  11. To view the EC50 or IC50 calculations, click on the individual curves (if you have more than one) and look for the C parameter listed above the dose response curve. In this case, this value represents the EC50 because this was an activation assay.
    EC50 value

    EC50 value

For more information or to download the free 14-day trial, please visit the MasterPlex ReaderFit product page.

Posted by aliu under MasterPlex ReaderFit

Join us for a Webinar on February 18
Space is limited.
Reserve your Webinar seat now at:
https://www1.gotomeeting.com/register/898296417
Please join us for our MasterPlex ReaderFit 101 webinar.  This webinar is designed for beginners and for people interested in seeing how the software works.  We will be going over the entire workflow of a sample analysis.  Topics include:


  1. Importing raw data
  2. Indicating sample type
  3. Entering standard information
  4. Acquiring the standard curve
  5. Applying quality control standards
  6. Viewing results
  7. Generating reports
  8. Creating templates
Title: Live FREE Webinar: MasterPlex ReaderFit 101
Date: Thursday, February 18, 2010
Time: 11:00 AM – 12:00 PM PST
After registering you will receive a confirmation email containing information about joining the Webinar.
System Requirements
PC-based attendees
Required: Windows® 2000, XP Home, XP Pro, 2003 Server, Vista
Macintosh®-based attendees
Required: Mac OS® X 10.4 (Tiger®) or newer

What’s New in this Upgrade?



For MasterPlex QT
  • Quality Control Manager (Optional)
    • Lower Limit of Detection (LLOD) – Automatically calculate your LLOD on a per analyte basis from the mean of the selected background wells plus the number of standard deviations you want defined
    • Percent Coefficient of Variation (% CV) – Define your maximum % CV threshold
    • Extrapolated Values – Automatically exclude extrapolated data from your results

  • Step-by-Step Wizard

    The new step-by-step wizard will guide you along the way from start to end.

  • Tooltip Popups for the Standard Curve

    Get at-a-glance views of all points on the standard curve by simply hovering over with your mouse.

  • Improved Best Fit

    The “Best Fit” feature has been improved to better pick the optimal model equation & weighting algorithm combination.
  • Copy & Paste Group Names

    You can now copy & paste group names just like with sample names.
  • Full Compatibility with FlexMAP 3D data formats
  • Full Compatibility with Multi-plate Batch Files
  • Full Compatibility with .XLSX Files
  • Full Compatibility with Windows XP, Vista, and 7

Download Free Trial Form


For MasterPlex ReaderFit
  • Quality Control Manager (Optional)
    • Lower Limit of Detection (LLOD) – Automatically calculate your LLOD on a per analyte basis from the mean of the selected background wells plus the number of standard deviations you want defined
    • Percent Coefficient of Variation (% CV) – Define your maximum % CV threshold
    • Extrapolated Values – Automatically exclude extrapolated data from your results

  • Step-by-Step Wizard

    The new step-by-step wizard will guide you along the way from start to end.

  • Tooltip Popups for the Standard Curve

    Get at-a-glance views of all points on the standard curve by simply hovering over with your mouse.

  • Improved Best Fit

    The “Best Fit” feature has been improved to better pick the optimal model equation & weighting algorithm combination.
  • Copy & Paste Group Names

    You can now copy & paste group names just like with sample names.
  • Full Compatibility with .XLSX Files
  • Full Compatibility with Windows XP, Vista, and 7

Download Free Trial Form


For MasterPlex EX
  • Step-by-Step Wizard

    The new step-by-step wizard will guide you along the way from start to end.

  • Copy & Paste Group Names

    You can now copy & paste group names just like with sample names.
  • Full Compatibility with .XLSX Files
  • Full Compatibility with Windows XP, Vista, and 7

Download Free Trial Form

The MasterPlex 21 CFR Part 11 Plug-in has just been released for use with MasterPlex QT v4.0, MasterPlex ReaderFit, and/or MasterPlex EX.

For organizations working under the FDA 21 CFR Part 11 regulations, this plug-in offers:

  • Multi-level user account management
  • Electronic Signature
  • Digital Signature on the digital outputs
  • File alteration detection
  • Log records
  • Audit trail (full log records)
  • Seamless connection with the xPONENT Luminex control software
  • A unique Finger Vein Identification option (PowerPoint Brochure, Product Page)

Download the FREE Trial version and request a license today!

The following webinar recordings have been made available for viewing at your own leisure:

Posted by aliu under MasterPlex QT, MasterPlex ReaderFit

We have a couple of free webinars coming up that I would like to announce:

  • 10 Tips for MasterPlex QT v4.0 Power Users on August 20th at 11AM – 12PM PDT – Reserve your seat!
  • Microplate Reader/Elisa Data Analysis made easy with MasterPlex ReaderFit on September 10 at 11:00 AM – 12:00 PM PDT – Reserve your seat!

If you cannot make the time for the MasterPlex ReaderFit webinar, you can view a previously recorded video of it and obtain the PowerPoint presentation slides.

A recording of the MasterPlex QT webinar will be available online after August 20th, 2009.

Posted by aliu under MasterPlex ReaderFit

The MasterPlex ReaderFit webinar covers the basic analysis of an elisa assay. Here is the list of topics that will be discussed in the recording:

  1. File Import Wizard
  2. Automatic Detection of Outliers
  3. Marking Wells and Grouping of Replicates
  4. Entering Standard Information
  5. Best Fit and Best Fit Parameters
  6. Plotting Unknown Data Points on the Standard Curve
  7. WYSIWYG Data Table
  8. Chart Customization
  9. Templates
  10. Custom Reports
  11. Virtual Plate

Download the PowerPoint Presentation if you would like to follow along with the recording.

Please click here to view the recorded MasterPlex ReaderFit webinar. Be sure to prepare some popcorn as the running time is about 40 minutes. Enjoy the show =)

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Posted by aliu under MasterPlex ReaderFit

Join us for our first webinar on using MasterPlex ReaderFit for microplate reader or elisa data analysis.

Join us for a Live FREE Webinar on August 6 2009 11:00 AM – 12:00 PM PDT:

Microplate Reader/Elisa Data Analysis made easy with MasterPlex ReaderFit






Space is limited. Reserve your Webinar seat now at:
https://www1.gotomeeting.com/register/364896864

We will be covering the following topics in detail:

  1. File Import Wizard
  2. Automatic Detection of Outliers
  3. Marking Wells and Grouping of Replicates
  4. Entering Standard Information
  5. Best Fit and Best Fit Parameters
  6. Plotting Unknown Data Points on the Standard Curve
  7. WYSIWYG Data Table
  8. Chart Customization
  9. Templates
  10. Custom Reports
  11. Virtual Plate

Here is a small MasterPlex Readerfit demonstration if you simply cannot wait.

We hope to see you there!

Posted by aliu under MasterPlex ReaderFit

The Camtasia Studio video content presented here requires JavaScript to be enabled and the latest version of the Macromedia Flash Player. If you are you using a browser with JavaScript disabled please enable it now. Otherwise, please update your version of the free Flash Player by downloading here.

The microplate analysis demo includes the following simple steps:

  1. Importing an Excel file with the RLU values
  2. Marking the background, standard, and unknown samples
  3. Filling in the standard concentrations with the AutoFill Feature
  4. Pressing the Best Fit button
  5. Reviewing standard curve, concentrations, and charts

Save time, stop copying & pasting, and download the FREE MasterPlex ReaderFit trial and see for yourself how easy it can be!

Posted by aliu under MasterPlex ReaderFit

The Hitachi Software MiraiBio group has just released a new build for MasterPlex ReaderFit, the quantitative analysis and curve-fitting data analysis software for microplate readers. New features and improvements include:

  • File Import Wizard – Although MasterPlex ReaderFit can automatically import some of the known file formats from common plate readers that are out there in the market, it does not handle all 100% of them. Unknown file formats can now be handled by the File Import Wizard in 1 of 2 ways:
    1. Plate Format

    2. List

    More details on using the File Import Wizard for MasterPlex ReaderFit

  • Best Fit Options – The Best Fit feature now offers 3 parameters for which you can optimize for to determine the best curve fit:
    1. Root Mean Square Error (RMSE) – RMSE is a good measure of accuracy or goodness of fit which aggregates the individual differences or residuals between your expected and calculated values into a single measure of predictive power. This is the default option as it is the one that is recommended.
    2. Standard Deviation of % Recovery – This option will provide the model equation and weighting algorithm combination with the lowest standard deviation of percent recovery. The percent recovery is basically the calculated concentration – the expected concentration * 100.
    3. R-squared – This option is the only Best Fit option in previous builds of MasterPlex ReaderFit. It is not as sensitive to the goodness of fit as the RMSE option.

  • Competitive Binding and Inhibition Assays – This build provides improved support for competitive binding or inhibition assays.

  • More Intuitive Auto Fill – The Auto Fill feature helps to automate the entry of the standard concentration values for serial dilutions. This feature has been simplified with more intuitive dilution direction buttons and being able to preview the standard values.

Try it out and download the free trial now!

Posted by aliu under MasterPlex QT, MasterPlex ReaderFit

The 5-PL or 5 Parameter Logistic is a nonlinear regression model used for prediction of the probability of occurrence of an event by fitting data to a logistic curve. It differs from the 4-PL or 4 Parameter Logistic model in that it is an asymmetric function which is a better fit for immunoassay or bioassay data.  As the name suggests, there are 5 parameters in the 5-PL model equation:

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