Best Practices for the FemtoQuest™ System with SMC® Technology
The FemtoQuest™ system, the next generation instrument beyond the SMCxPRO® immunoassay system, utilizes unique SMC® technology to provide high sensitivity measurement of your critical analytes. While the fluidics-free design allows for lower cost of maintenance and the compact footprint fits easily into lab spaces, this next generation instrument offers several advantages including dual plex and single analyte analysis, adaptive scanning that optimizes scan time (reducing variability in replicates), and automation and LIMS compatibility. Our best practices for using the FemtoQuest™ system include instrument maintenance guidelines, SMC® 384-well read plate handling, and general information regarding SMC® assays.
Section Overview
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FemtoQuest™ System Recommendations
The FemtoQuest™ system should be positioned in the lab at least 6 inches from the back wall to allow proper airflow behind the unit. It should not share a benchtop with any other equipment that shakes or causes vibration. The FemtoQuest™ instrument ensures thermal stability before beginning to read a plate. Establishing thermal stability can take from 90 seconds to 16 minutes.
The FemtoQuest™ system contains a Self-Test feature that verifies that the mechanical and optical components of your instrument are functioning normally. The Self-Test takes less than 5 minutes to complete and does not require a well plate to be present. Self-Test should be run daily and before the instrument is in use.
The SMC® Fluorescence Verification Kit is designed to ensure the FemtoQuest™ system is performing within normal parameters and consistently over time. The FemtoQuest™ system requires that the SMC® Fluorescence Verification Kit be run at least once per month. Failure to run the kit once every four weeks will prevent any well plate scans until the SMC® Fluorescence Verification Kit has been run.
How to Handle SMC® 384-Well Read Plates
SMC® assays are prepared in 96-well bottom Axygen plates until the end of the protocol, where eluted and neutralized samples will be transferred to SMC® 384-well Read Plates, provided in kits, for acquisition. The SMC® 384-well plates are the only compatible 384-well read plate for the FemtoQuest™ system. These plates also work in the SMCxPRO® instrument. These plates are extensively quality tested to provide optimal optical performance when handled correctly. Each plate includes a black lid which should be kept under the plate to protect well bottoms from scratches, fingerprints, dust, and debris. Cover the read plate wells immediately after removing them from packaging to minimize exposure to open air, lint, and dust.
Use only aluminum adhesive seals for assay plates that will be read on the FemtoQuest™ and SMCxPRO® instruments. Do NOT use clear acrylic seals to seal loaded read plates because they do not protect against light. The paint inside the instrument hood is reflective and will disrupt signal acquisition.
The SMC® 384-well Read Plates are labeled with a Radio Frequency Identification (RFID) tag and the FemtoQuest™ instrument is designed to work exclusively with these plates only. The type of assay you are running will determine the type of RFID-labeled well plate that is required.
Important: The number of analytes in the assay you are running must match the “plex” number of your RFID well plate. Always verify that you are using the correct RFID plate type before beginning your run.
Each RFID labeled well plate can be read a maximum of four times.
Figure 1.The aluminum adhesive seal that is needed for the 384-well assay plates read on FemtoQuest™ and SMCxPRO® instruments.
To ensure optimal acquisition precision, it is important to seal the read plate completely and uniformly to prevent well evaporation and cross-well condensation, which may occur if each well is not fully sealed.
To achieve complete sealing for each well:
- Use the recommended aluminum adhesive seals and plate roller
- Remove the backing of the seal and center the seal smoothly over all 384 wells without creating any creases, folds, or kinks
- Run fingers across the seal on the plate to secure placement, then create a complete seal by rolling the plate roller across the entire face of the plate:
- First, in a vertical direction
- Then, in a horizontal direction of the plate
- Hold the plate down with one hand. Tilting the plate roller on the plate with the other hand, use the soft corner of the silicone padding and migrate from one end of the plate to the other, vertically, then horizontally.
- Pay close attention to the junctions and corners of the outer edge wells and the plate frame.
- A complete seal will have a concave appearance across all wells, and all rims and well wall corners will be clearly visible.
Firmly and carefully place your loaded read plate containing sample into the Tray Load Portal, ensuring that all four corners are seated properly. If the plate is tilted during the well scan, data collection will be compromised, and the software may return an error.
In addition, flip up the perforated side strips of the aluminum adhesive seals before seating your plate into the instrument to prevent potential acquisition issues due to plate alignment.
Pulse centrifuge the 384-well read plates after each read to pull down any product condensation between reads. Carefully handle read plates to prevent disturbing the liquid contents.
Learn more tips in our “How to Seal Read Plates” article.
How do I avoid interference in signal detection?
Thorough elution and neutralization of immunoassay complexes and proper sample transfer to the SMC® read plate are essential to achieve stable, reproducible well RESPONSE signals and ensure intra-assay precision. Detailed neutralizations may be found within the kit protocol.
To avoid interferences in signal detection, avoid air bubbles when transferring the neutralized eluate to the 384-well read plate by aspirating and dispensing slowly using a P20 multichannel pipette. If bubbles persist, use air from a P1000 pipette to resolve them before applying the final read plate seal. Explore more pipetting tips in our "How to Properly Pipet” article.
Does the FemtoQuest™ system require a warm-up period?
No, however, the instrument will check thermal stability before beginning to read a plate. Establishing thermal stability can range from 90 seconds to 16 minutes.
Can SMC® assays be read multiple times?
SMC® assays can be read more than once. However, we do not recommend storing and re-reading the SMC® plates. Storage of plates at 4 °C followed by reading in the warm instrument can lead to evaporation and potential cross-contamination of wells. If an issue should occur during a run, the plate can be run again immediately without storing. For further guidance please call technical support.
Related Products
For Research Use Only. Not For Use In Diagnostic Procedures.
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