What can I use NuTec for?
NuTec is suited for developing highly diverse applications. Here are just a few examples.
- Diagnosing diseases in people and animals.
- Predicting who is likely to relapse following surgery or another treatment.
- Identifying novel biomarkers (i.e. small molecules or proteins).
- Detecting air or waterborne pathogens.
- Testing foods for the presence of pathogens.
- Wine/beer making.
- Identifying surfaces with specific properties: Anti-microbial, Cell supportive, Stem-cell differentiation, Drug binding for quantitation or nanoparticle design
In short, NuTec is limited only by your imagination. Note however that you may have to seek FDA approval for your particular application.
What sample types can I use with NuTec devices?
Just about any aqueous sample can be applied to NuTec devices. Prolonged exposure to strong acids and bases, and some organic solvents may destroy some surfaces on the device.
My application requires aseptic technique. Can I sterilize NuTec devices?
Yes, NuTec devices can be sterilized with gamma or UV radiation.
Can I culture cells on NuTec devices?
Yes, cell culture is easily performed on NuTec devices as follows.
- Place the sterilized NuTec in sterile water or culture media for 5~10 minutes to remove the glycerol preservative.
- Place the NuTec in a sterile container and add cells directly onto the surfaces.
How can I decrease the amount of abundant proteins in my serum or plasma samples?
Various protocols that remove abundant proteins from serum or plasma are compatible with NuTec. The protocol below is provided as an example.
- Add 100 uL of serum or plasma to a 1.5 mL tube.
- Add 300 uL of ice cold acetonitrile.
- Vortex briefly and incubate at -20°C for 60 min. Vortex 2~3 times during the 60 min incubation.
- Centrifuge at 6000 RPM for 1 min.
- Collect the supernatant and place into a fresh 1.5 mL tube. We typically collect the entire sample above the pellet (including the interphase). This results in ~360-400 uL of sample.
- Add 700 uL of 50 mM ammonium acetate to the collected supernatant.
- Vortex and incubate for ~5 min at room temperature.
- Centrifuge the mixture at 6000 RPM for ~10 seconds to pellet debris.
- Apply 1 mL of mixture onto the NuTec.
- Incubate sample on the NuTec for 60 minutes at room temperature.
Do samples need to be pre-processed before applying them onto the NuTec?
Most samples (e.g. urine, saliva, cell culture media, etc.) do not require pre-processing unless special considerations apply. Here are a few examples.
- Urine from diabetic patients have too much sugar and should be diluted 1:1 with 50 mM ammonium acetate to decrease the signal from sugar.
- Serum and plasma samples can be pre-processed to remove abundant proteins that can mask the detection of rarer molecules.
- Concentrated cellular or tissue lysates can be diluted with 50 mM ammonium acetate if needed.
Developing NuTec signatures
- Submerge NuTec in distilled water for 5~10 min to remove the glycerol preservative on the surfaces.
- Hold the NuTec firmly by the label, tap onto a paper towel and flick several times to remove excess water from the surface. Alternatively, blow filtered air over the NuTec to remove excess water from the surface. It is important not to let the surfaces dry.
- Place the NuTec on a flat table and add 1~2 mL of your sample over the surfaces. Make sure all surfaces are submerged in your sample. Use a consistent volume of sample through-out the study for each NuTec.
- Incubate at room temperature for 10~60 minutes. The incubation time depends on your sample type and the signal it produces. For reference:
- Urine, wine, milk, culture media and most samples require a 15~20 minute incubation period.
- 5% serum in 50 mM ammonium acetate requires 60 minute incubation period.
- Serum depleted of abundant proteins in 50 mM ammonium acetate requires 60 minute incubation period.
- Hold the NuTec firmly by the label, tap onto a paper towel and flick several times to remove excess sample from the surfaces. Alternatively, blow filtered air over the NuTec to remove excess sample from the surface.
- Heat NuTec to 300C for 5 minutes to develop the signal. A toaster oven that’s not pre-heated works well. A hot plate can also be used and heated for 3~5 minutes. The important thing is to use consistent conditions through-out the study.
- Allow the NuTec to cool for ~10 minutes before scanning.
Image acquisition using a Photo Scanner in Transmittance mode
- Any Photo Scanner with Transmittance mode (Epson Perfection V500 or other photo scanners) is appropriate for image acquisition. Transmittance mode means that the NuTec should be illuminated from the one side of the scanner and scanned on the opposite side.
- Fit the developed NuTec into the custom slide holder provided, then place the slide holder with the sample spots facing downwards towards the scanning bed. Take special care not to damage the developed surfaces.
- Before scanning, preview the image to be scanned and use the marquee function to select the area to be scanned. This are should include the NuTec surfaces and a few millimeters of the slide holder
- Scan the selected area in transmittance mode at 1200 dpi resolution.
Image acquisition using a Smart Phone
This scanning option is currently in development
How do I analyze my NuTec signature?
- Send scanned images to Entopsis (email@example.com). We are in the process of developing an online and automated solution for analyzing samples with NuTec. Please pardon the temporary inconvenience.
- The process will likely involve the following:
- Select one or several assays on the Entopsis website or app. You will get results from assays which you select.
- Upload the signature you want analyzed.
- The site will provide you with results and confidence indicators.
How do I make my own assay?
- Collect samples that are known to be positive and negative for the condition you want to test. For example, samples from breast cancer patients and people known to not have breast cancer.
- Try to use samples that are similar to each other, with the exception of the condition you want to test (e.g. age and gender matched, collected and stored in the same fashion, etc.) These samples make-up the data set that’s used to train the machine learning algorithm for your assay. Your assay is only as good as your training set.
- Obtain signatures from your labeled samples and perform image acquisition as described above.
- Send these images to Entopsis for assay development (firstname.lastname@example.org). We are in the process of creating an online and automated solution for assay development. Please pardon the temporary inconvenience.