A. General Questions
1. I need an in-person discussion or Zoom meeting to design my experiment for grant submission/manuscript writing. How can I start?
Contact the core via email (ORIONCore@mdanderson.org), and we will set up a time for you.
2. How do you provide services for clinical studies?
Most clinical studies can be set up through the Institutional Tissue Bank (ITB) to have fresh tissue or banked PBMC sent directly to the core. We also work with CTRC, MelCore and other biobanks on campus.
3. How do I access the data?
ORION Core uses Teams as a Sharepoint front-end for data upload. Upon completion you will be sent access to a personal folder for your project to download from.
4. Can I be trained to use instruments for unassisted use?
Training is available for the CTL ELISPOT analyser and MSD QuickPlex SQ120 only.
5. What platform are you using for cytokines and chemokines detection?
We have the Luminex and MSD platforms.
B. Luminex and MSD
1. Do you carry any Luminex/MSD kits at the core?
No, all kits are ordered at the time a request is made.
2. How to purchase kits Luminex/MSD kits: through the core or by ourselves?
We recommend purchasing through the core as we are able to leverage discounts, track receipt of the kit, and ensure proper storage. If you are an MD Anderson user, please provide the core with a valid chartfield by email for ordering. If you are non-MD Anderson user, the cost of the kit will be added as an ad-hoc charge in iLab.
3. What is the shelf-life of the Luminex/ MSD kits?
Most kits arrive with a shelf life of 1 year. We can verify expiration dates for you prior to ordering.
4. How much sample volume do I need for Luminex or MSD assays?
This can vary due to recommended dilution factors for a given kit/set of analytes or by sample type. But undiluted most kits require either 25µl per well of serum/plasma or 50µl per well of cell culture supernatant or lysate. We recommend running samples at minimum in duplicate.
5. What sample types/species are allowed for Luminex or MSD assays?
We can run most sample types and species. Common sample types are: serum, plasma, CSF, cell culture supernatants, tissue or cell lysates.
6. What is the workflow to get services for Luminex or MSD assays, and how long does it take to get data?
Luminex and MSD assays should be submitted through our iLab portal. We operate on a first-come, first-serve basis and schedule assays only after we receive the samples, kits, and iLab request. Once all items are in place, standard assays normally turn around in 1 to 2 weeks.
7. How do I prepare samples for Luminex/MSD?
Plasma: collect samples in sodium citrate, EDTA, or lithium heparin tubes (check what works best with the analytes you’ve selected). Centrifuge 1000xg at 4°C 10 mins within 30 mins of collection. Immediately store aliquots at -80°C.
Serum: allow blood to clot 20 mins at RT. Centrifuge 1000xg at RT 10 mins. Immediately store aliquots at -80°C.
Cell culture supernatants: centrifuge samples 1400rpm at 4°C 10mins to remove cellular debris. Aliquot into polypropylene tubes or plates and immediately store at -80°C. Please be prepared to provide 5mL of the cell culture media used in your assay, preferably in a serum-free format.
For all sample types, avoid freeze/thaw cycles.
8. How should I transfer my samples to the core?
If you are an MD Anderson user, we recommend using the Courier Services (Courier@MDAnderson.org) and requesting a STAT transfer before 3 p.m. Email ORION core for assistance with a courier form for transfer.
If you are a non-MD Anderson user, you can ship your samples to us or we can meet you outside of our building for pickup.
Samples should be transferred on plenty of dry ice, and an Excel manifest should be emailed to ORIONCore@mdanderson.org or uploaded to your iLab request. Please arrange your samples in the box in the order on the list.
9. How are the samples stored?
Upon receipt, Luminex and MSD samples are immediately stored at -80°C.
10. I’ve never performed a Luminex or MSD assay and I’m not entirely sure what I’m looking for. What’s a good place to start?
First, consider what proteins (analytes) you want to prioritize for your hypothesis. We can help you look for kits that encompass these and others in your area of interest. There are broad Luminex kits that cover many cytokines/chemokines/growth factors in up to an 80-plex that can be run on a single plate. MSD plates can only accommodate up to a 10-plex per plate. Please note that not all analytes can be plexed together due to cross-reactivity or differing sample dilution needs.
11. What is the minimum acceptable bead count for Luminex assays?
35 beads per analyte.
C. Flow Cytometry
1. What sample types/species are allowed for flow cytometry?
We only accept human samples for flow cytometry. We can analyze PBMCs, bone marrow and fresh tissues.
2. Do you provide unassisted use (self-use)?
We do not. Please see the MD Anderson north campus Flow Cytometry and Cellular Imaging Core Facility or the south campus Advanced Cytometry & Sorting Facility for these services.
1. Can I make my own plate and bring it to read myself?
Yes, you can request a training session in iLab under the Request Services tab or by emailing the core. Once trained, you can reserve the instrument in iLab under the Schedule Equipment tab. The CTL ELISPOT reader can only be scheduled during core hours, from 10 a.m. until 6 p.m.
2. I don’t have time to make a plate. Can you do everything for me?
ORION can perform a colorimetric IFN-gamma peptide ELISPOT assay on human cells using either short or long peptides. All other assay types must be performed by the user at this time and can be brought to the core for scanning/counting.
3. When can I use the ELISPOT reader?
The CTL ELISPOT reader can only be scheduled during core hours, 10 a.m.–6 p.m.
4. How do I reserve the CTL ELISPOT reader?
Once trained, you can reserve the instrument in iLab under the Schedule Equipment tab.
5. How do I get my data transferred?
Assays completed by the core will be uploaded into a private Teams folder for download. Users analyzing their own plates may log in to their own OneDrive or Teams folders to upload.
6. Where is the ELISPOT reader located?
It is located by the lab bay 2SCR2.3219. An ORION core staff member will log you in at the time of your arrival. Please enter through the rear lab door where you will see ORION signage and a doorbell.
7. Who do I “check in” with? How do I contact them?
We will arrange to have one of our core staff in the lab at the time of your reservation. You can also contact us by emailing ORIONCore@mdanderson.org or calling us at 713-563-1955.
8. Will you help me if I have a problem?
Yes, we will be able to help with login and general instrument issues. If an issue occurs that requires senior staff, such as difficulty analyzing/interpreting a plate with a high background or unusual spots, we may have to arrange a separate time slot for plate review based on their availability.
9. What happens if I need more time?
Once we verify no further users are expected after you, you can use the instrument until 6 p.m. and we can adjust your usage time in iLab.