Hitachi Solutions America, Ltd.
MiraiBio Group
Analysis Software for the Life Sciences
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Importing data into MasterPlex ReaderFit

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With the large number of plate readers and/or file formats out there it would be very difficult for us to have native support for each one in MasterPlex ReaderFit. That being said we wanted to make data import as easy as possible so ReaderFit allows you to import your data in multiple ways:

1. Copying and Pasting data into ReaderFit

  • Simply copy your raw data. It can be in plate shape or list form.
  • Left click to select well A1 then Right-click on it, select Paste Data and then Paste Response Values:

2. Using the File Import Wizard

-There are 2 supported formats for the File Import Wizard:

  • Plate - This format is in the 96-well plate format where all the response or raw values are in a table that corresponds to all 96 wells.

Here is the procedure for handling Plate format:

1. Deliminator Type - Select the deliminator used to separate all the values in your file

2. Data Format - Select the Plate option

3. Select Data - Click and drag to select the area in the plate view window on the left where all the values are located and click Next

4. For each analyte set that you have (not common), highlight and select the corresponding wells and click Assign

5. Click Finish

  • List - This format is a 2-column format that indicates well location and the response or raw value. (Note: in later versions support for a 3rd column containing sample name was added.)

Here is the procedure for handling List format:

1. Deliminator Type - Select the deliminator used to separate all the values in your file

2. Data Format - Select the List option

3. Well Address Selection - Highlight and select all the address wells in your list file in the preview window on the left and click Done for Well Address. Please note that the only accepted well address formats accepted are the conventional addresses for 96-well plates such as “H3” or just plain numbers.

 

4. Data Selection - Highlight and select all the data wells in your list file in the preview window on the left and click Done for Intensity.

5.Optional: If you have Sample Names you can follow the same procedure as in steps 3 and 4 to import sample names.

6. Click Next

7. For each analyte set that you have, highlight and select the corresponding wells and click Assign

8. Click Finish

Copying your raw data from an Excel sheet or text document and pasting it directly into ReaderFit is highly recommended as it is the fastest and easiest method for importing data!

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How to install a Network License in MasterPlex

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This procedure is for the Network license installations only.

MasterPlex Network License Manager Software Installation

  1. If you have not done so yet, download the MasterPlex Network License Manager.
  2. During this installation, please make sure you are logged on as administrator or
    have administrative privileges.
  3. Please make sure that the license server computer is connected to the local area
    network. It is required for proper function of the software.
  4. Please launch MasterPlex Network License Manager 2.0 downloaded from step1 and the InstallShield Wizard will guide you through the installation.

Start the MasterPlex Network License Manager Service

  1. On the Desktop, right-click on the My Computer icon and select Manage.
  2. Double-click Services and Applications and then Services to show the list of
    services.
  3. Right-click on MasterPlex Network License Manager and select Start.Image
  4. Make sure the service status has changed to Started.

Obtain Network License for MasterPlex Software

  1. Launch the MasterPlex Network License Manager and click on the Obtain License button. Press Unblock if the following window appears:
    Image
  2. Select MasterPlex License Manager from the pull-down menu and click Proceed
  3. Please note the Machine ID in this window as it is specific to the computer and is required when requesting for your license file.
  4. Next, please note the Validation Code that is included in the documentation or email when you purchased your software. If you made your purchase online, you will most likely receive your Validation Code via email. Otherwise, your Validation Code will be mailed to you along with the software package.
  5. Once you have your Machine ID and Validation Code squared away, please use our Product Registration Form to request for your license file.
    • License type – Select the Network radio button.
    • # of Concurrent Users – Choose the number of concurrent users based on the number of Validation Codes you have purchased. You will only have to specify 1 Validation Code because they will all be linked in our system
  6. Press the Submit button once you are done and you will receive your license file via email within 1 business day.

Install Network License for MasterPlex Software

  1. Once you receive your license file, save it to your local computer.
  2. Make sure you have Administrative Privileges before proceeding because it will be required for the successful installation of your license file. If you are unsure of the privileges to your user account, please consult with your IT group.
  3. Launch the MasterPlex License Manager.
  4. Click on the Install License button.
  5. Browse to your .lic license file and press Open.
  6. You should now be able to see the license you have just installed in the Installed Licenses table.
    ImageIf the installation is successful, you will see a message saying that the license was successfully installed. If you run into an invalid license error message, please see the following list of some common causes:

    1. Make sure that you have administrative privileges when installing the license. You may need to contact your IT department to ask for assistance with this.
    2. Make sure that the Machine ID originated from the same computer that contains the MasterPlex Network License Manager and not one of the client computers. Also, double check that the machine ID is correct. Note that the “0″ may look like an “O” but there is no letter “O” in the machine ID. Please resubmit for another license if you have reported an incorrect machine ID.
    3. Make sure you have submitted the request for the correct software version. Please resubmit for another license if you have reported an incorrect version number.

Launch Client Software

  1. When an unlicensed version of MasterPlex is initially launched, a user will see the License Information window. If the License Information window does not pop up, you can do the following:
    • For MasterPlex QT v5.x, MasterPlex ReaderFit v2.x, and MasterPlex EX v2.x, go to License->Applications and click on the splashscreen for the application of interest.
    • For MasterPlex QT v4.x, MasterPlex ReaderFit v1.x, and MasterPlex EX v1.x, go to Help/Tools->Applications and click on the splashscreen for the application of interest.Image
    • For MasterPlex CT, MasterPlex GT v2.3 and below, MasterPlex QT v3.0 and below, and MasterPlex QT EX, go to Help->License Information.
      ImageImage
  2. Select Network Client License Mode and click Auto Detect to find the MasterPlex Network License Manager automatically.
  3. If it cannot find the license server automatically, you can manually type in the server information (IP address and port number) and click the Test button.
    Image
  4. To launch the software, press OK.

If you run into any issues launching or finding the MasterPlex Network License Manager, please make sure of the following:

  • The MasterPlex Network License Manager service has been started. (Please see instructions above for Starting the MasterPlex Network License Manager Service.
  • If you are using a firewall, please make sure the following ports have been added to the exceptions list:
    • 12405 (TCP)
    • 12483 (UDP)

    If you have edited the port numbers, please make sure you use your configuration instead.

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How to install your license of MasterPlex or ReaderFit

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MasterPlex software has a 14-day temporary license when it is first installed. After this time period has passed a user will have to install a product license to continue to use the software. When an unlicensed version of MasterPlex or ReaderFit is initially launched, a user will see the License Information window.

  1. Please note the Machine ID in this window is specific to the computer and is required when requesting your license file. If the License Information window does not automatically pop up, you can do the following:
    • For MasterPlex GT v2.3 and below, MasterPlex QT v3.0 and below, and MasterPlex QT EX, go to Help->License Information.
      ImageImage
    • For MasterPlex QT v4.x, MasterPlex EX v1.x or MasterPlex ReaderFit v 1.x go to License->Applications and click on the splashscreen for the application of interest. (This step is important. There are 3 different modules in the MasterPlex suite and their icons are tiled in the License Information box. Make sure you select the correct module.)Image
    • For MasterPlex QT v5.x, MasterPlex EX v2.x or MasterPlex ReaderFit v 2.x go to License->Applications and click on the splashscreen for the application of interest. (This step is important. There are 3 different modules in the MasterPlex suite and their icons are tiled in the License Information box. Make sure you select the correct module.)

    License Information Window
    Image

  2. Next, please note the Validation Code that was included in the email when you purchased your software.
  3. Once you have your Machine ID and Validation Code squared away, please use our Product Registration Form to request for your license file. (Note: For License type, make sure you click the Term radio button.)
  4. Press the Submit button once you are done and you will receive your license file via email within 1 business day.
  5. Once you receive your license file, save it to your local computer.
  6. Launch the MasterPlex software to access the License Information window again.
  7. Press the Install License File button and browse to the location of your file and press Open.

If the installation is successful, you will see a message saying that the license was successfully installed. If you run into an invalid license error message, please see the following list of some common causes:

  1. Some older versions of MasterPlex require administrative privileges when installing the license. You may need to contact your IT department to ask for assistance with this.
  2. Make sure that the Machine ID is the same ID as the one you have submitted for the license file. Also, double check that the machine ID is correct. Note that the “0″ (zero) may look like an “O” but there is no letter “O” in the machine ID. Please resubmit for another license if you have reported an incorrect machine ID.
  3. Make sure you have submitted the request for the correct software version. Please resubmit for another license if you have reported an incorrect version number.
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How to export standard curve charts in MasterPlex ReaderFit and MasterPlex QT

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There are several ways of exporting standard curve charts in MasterPlex and they are all available under the “Standard Curve” or “Fit Curves” tab:

  1. If you are only interested in exporting a particular standard curve. Make sure it is the active chart and simply right-click on the chart to pull up some export options.Image

    You will then have the option to either copy the image onto a clipboard to paste into a document or presentation or export the image into many different file types such as Bitmap, PNG, HTML, PDF, or Excel.

  2. If you want to export all the standard curves in one report, then click on the Print button Image. Next, select the All radio button and press OK. Once the Preview window pops up, click the Export Document button and numerous export options will appear that include:
    • PDF
    • HTML
    • MHT
    • RTF
    • Excel
    • CSV
    • Text
    • Image

    Image

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How Not to Draw Standard Curves

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  • Using a linear curve fit (or regression) in Excel (y = mx + c)

    • Excel is an extremely powerful tool designed for a wide range of applications but drawing standard curves and interpolating unknown concentrations for ELISAs assays is not one of them. Bioassays or immunoassays resulting from biological systems follow a trend that is more analogous to the four paramater logistic (4PL) or five parameter parameter logistic (5PL) models rather than a linear model because there are limits to every biological system whereas linear curve fits go to infinity.
      4 Parameter Logistic Model

      Sigmoidal 4PL Curve Fit

    • Simply plugging in 0.5 for y and solving for x using a linear equation will get you the midpoint of the line but it is not an accurate IC50 or EC50 value because we are starting off with a very crude equation or approximation of the true curve fit. Simply put, you are forcing a square peg into a round hole. The more accurate method of obtaining the IC50 or EC50 value would be to use the 4PL model equation to do the curve fit and using the calculated C parameter.
      C Parameter - Inflection Point of the curve that also represents the EC50 or IC50

      C Parameter – Inflection Point of the curve that also represents the EC50 or IC50

  • Using the trendline feature with a scatter plot in Excel to get the equation and parameter values

    • Excel only offers linear, logarithmic, polynomial, power, exponential, and moving average as options for the trendline (reference). None of these are sufficient enough to describe the sigmoidal shape of immuno and bioassays.
  • Drawing standard curves without using weighting to offset heteroscedasticity

    • Heteroscedasticity is a phenomenon in which the variance of the dependent variable varies across the data. When dealing with [MFI/absorbance values/RLU] and concentration values, the concentrations usually increase as the [MFI/absorbance values/RLU] increases. When dealing with the high end of the standard curve, it is natural for the concentration values to have a greater variance when compared to the small concentration values on the low end of the standard curve. The use of weighting algorithms such as 1/y or 1/y^2 can be used to deal with the variance. More details can be found here.

The correct way of doing 4PL curve fits in Excel usually involve having to install an add-on that can do 4PL curve fitting but not all add-ons support weighting.

ReaderFit provides 4PL and 5PL curve fits with 4 different weighting algorithms.

ReaderFit.com – Free online curve-fitting application
Sign Up for Free Account

ReaderFit Desktop – Robust curve-fitting, quality control and reporting desktop software
Download Free Trial

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What to do when ReaderFit or MasterPlex launches with error on non-English Windows OS

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MasterPlex software is used by 1000′s of people across the world to analyze single and multi-plex ELISA data. Not everyone is running the English version of Windows! Here is some information to help users on non-English Windows installations who are running into problems installing and using MasterPlex software.

Symptoms:

  1. MasterPlex crashes when trying to start.
  2. User is running on non-English version of Microsoft Windows
  3. You see an error message such as this: MasterPlex is closed due to one or more errors. Le format de la chaine d’entree est incorrect.
  4. You use a decimal comma in your numbering system.
  5. You see an error message such as this: MasterPlex is closed due to the following error: Invalid number format.

Cause:

MasterPlex’s numbering system is set to use the comma “,” as the number grouping symbol and the period “.” as the decimal indicator symbol. Any difference in the numbering systems, for example the Italian numbering system, will cause MasterPlex to crash.

This problem can be solved by choosing English as the Standards and formats from the Regional and Language Options control panel.

Solution:

Change the Regional Language settings in Windows to correspond with the numbering settings needed by MasterPlex.

Windows XP

  1. Go to Start > Control Panel.
  2. Click on Regional and Language Options.
  3. Go to Regional Options tab > Standards and formats section and change option to “English (United States)”.
  4. Click OK to confirm the changes.
  5. The computer may ask you to restart, please do so if asked.

Windows Vista

  1. Go to Start > Control Panel.
  2. Click on Regional and Language Options.
  3. Go to Formats tab > Current Format section and change option to “English (United States)”.
  4. Go to Location tab > Current Location and change option to “United States”.
  5. Click OK to confirm the changes.
  6. The computer may ask you to restart, please do so if asked.

Windows 7

  1. Go to Start > Control Panel.
  2. Click on Region and Language.
  3. Go to Formats tab.
  4. Change to “English (United States)”.
  5. Click OK to confirm the changes.
  6. The computer may ask you to restart, please do so if asked.

Known Affected Versions:

  • MasterPlex QT: 4.x, v2.0
  • MasterPlex ReaderFit: 1.x, v2.0
  • MasterPlex EX: 1.x, v2.0

Don’ have MasterPlex or ReaderFit installed yet?

ReaderFit Desktop – Robust curve-fitting, quality control and reporting desktop software for single plex ELISA data (all Plate Readers)
Download Free Trial

MasterPlex QT – Robust curve-fitting, quality control and reporting desktop software for multiplex ELISA data (Luminex, Bio-Plex, Meso Scale Discovery and Applied BioCode platforms)
Download Free Trial

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Top 5 reasons why Luminex/Bio-Plex® users prefer MasterPlex® analysis and reporting software

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What is MasterPlex?

 MasterPlex is a flexible quality control, curve fitting, graphing and reporting software package for xMAP® quantitative data sets. The MiraiBio group of HITACHI has been offering MasterPlex software as a value-add to the Luminex xMAP and Bio-Rad Bio-Plex systems for 10 years.

 Why would I use MasterPlex?

 1.      You like to tweak your curve fitting settings: MasterPlex is great for labs that want to standardize on one curve fitting model equation (see the section on templates below) but it does really well for users that like to get their “hands dirty”. In fact, MasterPlex has a whole tab dedicated to curve fitting. A user can instantly toggle between fitting curves to the average value of replicates or treat each replicate as an individual point. Any combination of 6 different model equations and 4 optional weighting algorithms can be applied on a per analyte basis or you can choose a custom fit to apply to the entire data set. As a bonus, use the BestFit feature and let MasterPlex choose for you!

 2.      You have advanced reporting or exporting needs: MasterPlex has a very cool drag and drop report builder right in the software. Easily add or remove one of the 21 different available data types to the report. Drill down, Excel-style into your data. Export to PDF, Text, Excel, CSV or HTML. The best part is that you only have to create a custom layout once. Save it in the template manager and reuse it with 1 click!

 For even more advanced reporting needs MasterPlex has an Export Manager that supports multi-sheet Excel reports and various text and XML exporting formats that are often used to import data into a LIMS or data base.

 3.      You want to save time on your analyses: MasterPlex utilizes reusable templates to automate many steps in the analysis process including indicating QC and analysis settings and applying custom report formats. Even a novice user utilizing the template managers can move from data import to final report export in less than 10 seconds!

 4.       You want to standardize on one software package for all of your quantitative data: Most labs are generating quantitative data from multiple platforms. MasterPlex reads data from the following sources:

·         LX/Bio-Plex 100

·         LX/Bio-Plex 200

·         FlexMAP 3D

·         MAGPIX

·         Meso Scale Discovery SECTOR®

·         BioCode

·         Most plate readers

Imagine standardizing not only on one analysis software package but on one report format!

 5.       You want to simplify your training process: MasterPlex has an intuitive tabbed interface that is supported by a number of short, easy to digest videos that can walk any user through analysis. Many labs like the fact that when a new member joins the team it does not take more than 30 minutes to get them trained on MasterPlex. Rather than having to train someone on multiple pieces of analysis software, many MasterPlex labs only have to train once.

 How do I learn more?

 Download a free fully functional 14-day trial today.

 

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InspireTheGenome.com Survey Winners Announced

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Thank you to everyone who took the time to fill out our survey at InspireTheGenome.com!

The following ten randomly selected winners received $25 gift certificates from Amazon.com:

Ying Lin
Sayed Mostafa Hosseini
Wenjun Li
Jie Wang
Andrea Semler
Dr. Nirankushk Khubalkar
Wojciech Rychlik
Osama Youssef
Gerardo Sanchez
Andy Michel

If you haven’t done so already please stop by InspireTheGenome.com to see HITACHI’s genomic data infrastructure concept. The survey link is still active too!

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HITACHI to show a sneak peek of a new cloud genome infrastructure at ASHG conference November 7-9, 2012

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Hitachi Solutions and Hitachi High Technologies have partnered to leverage breakthrough technologies to create the secure infrastructure needed to allow validated genomic data and related phenotypic data to be combined. This new platform will give users the ability to mine and view data in novel ways to make new discoveries.

Additionally, the platform is built on the paradigm that the individual, not health care or insurance providers, controls their genomic data and who may have access to it. The concept has tools to allow users to grant access to their personal health information, including genomic data, to specific providers.

That is not to say this concept is solely another entrant in the increasingly crowded personal health data space. Rather the platform also leverages HITACHI’s strong background in security, coupled with their unique technology to anoymize user data. This will offer health care, application and content providers access to incredibly valuable data, such as population studies, without the traditional risks involved in compiling and studying this type of data.

Please stop by HITACHI’s booth (#1502) at the American Society of Human Genetics Conference at the Moscone Center in San Francisco Nov. 7-9, 2012 to see a demo and learn more.

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