STarStation FAQ's

Software:

1. Which version of Luminex software is compatible with STarStation 2.3?

2. Will I lose my data if an acquisition is stopped during a run?

3. I have finished my acquisition and noticed I have entered a bead ID incorrectly. Is there any way to amend this?

4. What is the default password for the admin log in account?

5. Is there a tutorial available for the STarStation software?

6. Where is the worklist .xml file saved to?

7. Where can I find my results after acquisition?

8. Can I export my results to excel?

9. Can I save on reagents and apply data of a standard curve from one plate to data of a sample only plate?

10. Can I use STarStation for SNP analysis?

11. What is the minimum number of microspheres that must be counted for each region?

Website:

1. Where can I find calibration and control target values?

2. Where can I find sample protocols?

Reagents & Consumables:

1. What is the diameter of the microspheres?

2. Approximately how much sheath fluid is used for a 96 well plate?

3. What is the shelf life of the sheath fluid?

4. How many microspheres are in a 1 ml stock vial?

5. What is the shelf life of the Luminex microspheres?

6. How many carboxyl groups are on the xMAP microspheres?

7. How should the microspheres be stored?

8. Can oligonucleotide-coupled microspheres be frozen?

9. Is it possible to store completed assays overnight prior to analysis?

Instrument:

1. How often should I replace the sheath filter on the Luminex analyzer?

2. What is the sensitivity of the Luminex instrument?

3. What are the specifications for the Luminex 200?

4. How long does it take to read a 96 well microtitre plate?

STarStation Statistics:

1. What does the R-squared value of the standard curve signify?

2. How are the various MFI calculations obtained?

3. What does Bead CV% mean?

4. How is the mean concentration determined?

5. How is the CV% (n) calculated?

Software

1. Which version of Luminex software is compatible with STarStation 2.3?

STarStation 2.3 is compatible with Luminex IS 2.3 software. STarStation 2.0 is compatible with all versions of Luminex software earlier than IS 2.3.

2. Will I lose my data if an acquisition is stopped during a run?

No, STarStation saves data in real time after reading each well into the samples folder of the specific user directory.

3. I have finished my acquisition and noticed I have entered a bead ID incorrectly. Is there any way to amend this?

Yes, this error and any others relating to the worklist such as standards or plate layout can be amended using the Worklist Editor, which is an additional piece of software. Please contact customersupport@appliedcytometry.com for more information.

4. What is the default password for the admin log in account?

The default password is admin.

5. Is there a tutorial available for the STarStation software?

Yes, the user manual is available online in the Guides section under Tutorial.

6. Where is the worklist .xml file saved to?

It can be found in the worklist folder of the user that created it.

7. Where can I find my results after acquisition?

These can be found in the samples folder of the user that was logged in at the time of acquisition.

8. Can I export my results to excel?

Yes you can do this by following the instructions that are found in the Guides section under Tec Bulletins. By using a simple copy/paste function, standard curves can be added to the excel report also. (create link to Guides section)

9. Can I save on reagents and apply data of a standard curve from one plate to data of a sample only plate?

Yes this can be achieved by using an additional piece of software called STarCollate. It allows you to quantify data from up to 20 previously acquired assay plates from the same standard curve. For more information, please contact customersupport@appliedcytometry.com.

10. Can I use STarStation for SNP analysis?

Yes STarStation can be used to acquire the data from SNP assays, then the data can be analysed by using an additional piece of software called STarBase. Click HERE [swf flash file] for demo.

11. What is the minimum number of microspheres that must be counted for each region?

Depending on the precision that you require for your assay, anywhere between 30 and 100 microspheres is sufficient. More microspheres counted better precision. Commercial cytokine assays suggest 100 microspheres.

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Website

1. Where can I find calibration and control target values?

These can be found in the Technical Support under Related Links.

2. Where can I find sample protocols?

These can be found in the Technical Support under Related Links.

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Reagents & Consumables

1. What is the diameter of the microspheres?

The microspheres are 5.6µm in diameter

2. Approximately how much sheath fluid is used for a 96 well plate?

About 1 litre of sheath fluid is used.

3. What is the shelf life of the sheath fluid?

For the 1x sheath fluid, the shelf life is 2 years from manufacturing date. Luminex does not ship out any sheath fluid with less than 6 months remaining. For the 20x sheath fluid, the shelf life is 9 months undiluted, 3 months diluted.

4. How many microspheres are in a 1 ml stock vial?

12.5 million COOH microspheres/ml 2.5 million LumAvidin microspheres/ml 250,000 FlexMAP microspheres/ml

5. What is the shelf life of the Luminex microspheres?

FlexMAP- frozen 6mths, 2-8°C 3mths Carboxylated- 3yr LumAvidin- 6mths

6. How many carboxyl groups are on the xMAP microspheres?

The number of carboxyl groups found is 1.10E+8

7. How should the microspheres be stored?

The microspheres should be kept at 2-8°C and in the dark. Avoid glass front refrigerators, organic solvents and freezing.

8. Can oligonucleotide-coupled microspheres be frozen?

Oligonucleotide-coupled microspheres are stable through one freeze-thaw cycle.

9. Is it possible to store completed assays overnight prior to analysis?

Provided the assay plate is kept in the dark, i.e. wrapped in tin foil, and at room temperature or 4°C, then it can be analysed the next day.

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Instrument

1. How often should I replace the sheath filter on the Luminex analyzer?

This should be replaced once a year and it is part of the annual PMI visit.

2. What is the sensitivity of the Luminex instrument?

From the aspect of assays, the instrument will only be as sensitive as you reagents. Commercial kits can measure concentrations as low as 0.1 pg/ml. The instrument itself can detect as few as 100 fluorescent phycoerythrin molecules per microsphere and can report signals as high as 32,767 median fluorescent intensities.

3. What are the specifications for the Luminex 200?

Download a pdf document of the specifications HERE [PDF]

4. How long does it take to read a 96 well microtitre plate?

It takes somewhere 35-60 minutes depending on the assay set up (bead concentration per assay, total assay volume). Using a 100 plex assay, 9600 data points can be measured in 1 hour.

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STarStation Statistics

1. What doe the R-squared value of the standard curve signify?

The R-squared value signifies the goodness of fit for your standard curve and is measured between 0 and 1. When R-squared equals 1, all points lie exactly on the standard curve.

2. How are the various MFI calculations obtained?

Raw MFI- Median Fluorescence Intensity, no pre-processing is performed. Net MFI (Blank) - MFI values obtained from (Raw MFI-Blank MFI) Net MFI (Zero Standard) - MFI values obtained from (Raw MFI-Zero Standard MFI)

3. What does Bead CV% mean?

It is the coefficient of variation of the reporter fluorescence detected on the surface of the classified assay microspheres performing the test in the specified well.

4. How is the mean concentration determined?

It is determined as the mean of the calculated concentrations for each of the individual replicates.

5. How is the CV% (n) calculated?

It is calculated from individual replicate concentrations with (n) being the number of replicates.

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