PanCrisp Cartridge Holder (5-Column)

The main intention behind the design of the PanCrisp cartridge is to combine several different parts of sample processing into one unit. It is composed of five vertical columns arranged side by side in a single cartridge. Each column has a designated area for a particular component of the diagnostic process. The upper end of the columns provides a place to introduce a sample or reagents into the system, whereas the lower ends include the processing and detection areas. Such an arrangement reduces the need for transferring samples between separate tubes and prevents contamination during the diagnostic process. The first stage of the diagnostic process starts with introducing saliva into the designated area of the cartridge where samples should be located. Lysis buffer is used in the system to facilitate exosomes' destruction and the release of miRNA. The sample moves through the filtration region of the cartridge after lysis. The purpose of the filter is to hold unnecessary material and let desired molecular components pass through it. Additionally, the sample-processing area of the cartridge can include DNase that will reduce unnecessary DNA before the downstream detection reaction. After sample processing, the RNA is sent to the detection area containing the CRISPR-Cas13 reaction. Our PanCRISP system is capable of detecting multiple target miRNAs (miRNA-21-5p, miRNA-23a, miRNA-23b, miRNA-29c, miRNA-216). If the target sequence is present in the sample, the CRISPR-Cas13 complex is activated and cuts fluorescent reporter molecules. As a result, the fluorescence signal is produced which can be detected at the lower end of the cartridge. PanCRISP cartridge design does not require any additional manual transfer of the processed sample into another reaction tube. Cartridge Holder In order to keep these five columns together and steady during the tests, we have created a special cartridge holder, made via 3D printing. Our holder is supposed to ensure proper spacing of each column and their vertical alignment during sample processing. We can easily change and print such holders using an ordinary 3D printer and cheap PLA filament. Such 3D-printed parts are used in other low-cost diagnostic hardware designs in previous research as well. The lower part of our holder is designed based on the region of fluorescence detection in order to keep the sample at one place while measuring its fluorescence. Additionally, the holder can be altered in accordance with the tube size, filters or the volume of reaction in case we need to test the PanCrisp cartridge further. The inside will contain all of the tubes, with cartridges and contents, and the outside is meant to be like a box with 5 doors for fluorescence detection, and have a handle on top for easy handling for customers.

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