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Point of Care

 

For the detection and analysis of cells for POC applications, VyCAP has developed a disposable cartridge that contains a MicrosieveTM. The cartridge collects, stains and presents cells from sample fluids such that :

1.     Large volume (1 10 ml) bodily fluid samples can easily pass the microsieve without the use of external pressure

2.     Cells are collected and concentrated on the microsieve membrane

3.     The cells are brought into contact with the staining reagents that are present inside the cartridge and are next incubated with these reagents

4.     The microsieve is flat and non-fluorescent and fluorescence images of the collected cells are acquired

5.     Collected and stained cells are easy to count and to differentiate using fluorescence microscopy technologies

 

 

POC Platform Technology

 

Simple POC Cartridge

For sample volumes ranging from 100 microliters to 10 milliliters with low number of cells (1 100 per microliter) VyCAP designed a disposable cartridge that captures the cells and allows staining using your own staining protocols. The filtration action can be started and stopped by rotating the sample cup. The flow rate is depending on your sample and is maximal 1 milliliter / minute.

 

The microsieve is positioned at the bottom of the sample cup. The sample fluid is added to the cup. The fluid passes the sieve and cells stay behind on the sieve. Next the filtration is stopped and staining reagents are added onto the sieve. After incubation the filtration is started again. This can be repeated multiple times without losing the cells. Operation of the cartridge can be viewed here.

After completion of the labelling images of the collected cells are acquired using a fluorescence microscope.

 

 

 

All In One Cartridge.

Image below presents an overview of the POC platform. The POC cartridge is depicted in the center of the image. The cartridge contains a cup that can hold 100 microliter-10 milliliter of fluid sample. The microsieve is placed at the bottom of this cup. The cartridge is prefilled with reagents which are directed towards the collected cells after the filtration step is completed. After the cells have been stained the whole cartridge is transferred to a dedicated reader which automatically differentiates the collected cells by means of their fluorescence color and intensity. Depending on the application the microsieve design, number of reagents, number of washing steps, and sample volume can be designed to the needs. The operation of the POC cartridge in real time can be viewed here.

 

Image below presents cells that have been filtered from a Cerebrospinal fluid sample and are next stained using the POC cartridge as presented above. The microsieve membrane has a total surface of 3 x 3 mm2 and contains pores with a diameter of 2 microns. After the staining is completed fluorescence images of the membrane surface are acquired. The intensity of each cell is determined using image analysis routines and next the cells are differentiated based on their intensities.

In adults leucocytes levels of < 5 per l are considered to be normal. The POC cartridge is able to process volumes of up to 10 ml. For Cerebrospinal fluid typically a sample volume of 500 l is used which results in 2500 leucocytes at the threshold level of 5 per l. This assures a small statistical error at this low level.