In situ sequencing in single cells and tissue

Single cell sequencing (often referred to as fourth-generation sequencing) provides higher resolution of cellular differences and better understanding of the function of an individual cell in the context of its microenvironment. Several assays have been developed and partially automated systems for single cell genomics are available. But current technologies lack high throughput, especially for tissue samples. This is why faster systems are needed to make single cell genomics attractive and feasible for a wider variety of different applications. 

Model Theta provides a fast and reliable way to sequence cells and tissue with your preferred fluorescence in situ sequencing assay in a fast and reliable manner. The improved 3D imaging of Model Theta delivers particular benefits in terms of avoiding background fluorescence and separating fluorescent objects located in close proximity to each other.


Digital ELISA

The ability to detect antigens is commonly achieved by traditional ELISA. However, the sensitivity of ELISA is limited, which makes low-abundance antigens is difficult to trace and quantify. To accelerate the discovery and use of more sensitive diagnostic biomarkers, single molecule detection is crucial. In single molecule detection the individual antigens are digitally counted, in contrast to bulk measurements where the total fluorescence intensity is measured. Bulk measurements in general suffer at low concentrations due to high noise levels in the detectors and, for example, because of variations in droplet size.

In contrast to the few other commercial systems for digital ELISA, Model Theta doesn't require non-conventional complex assays and allows even higher sensitivity, far below fg/ml antigen concentrations and with a dynamic range of six orders of magnitude. Throughput is also heavily improved, which is particularly important when large patient samples need to be processed.


Cytology and digital pathology

Slide scanners and flow cytometry are routinely used in the diagnosis of health disorders. Recent advances in whole-slide imaging have resulted in digital pathology being seen as one of the most promising avenues in diagnostic medicine for achieving better, faster and cheaper diagnosis, prognosis and prediction of cancer and other important diseases. But most instruments still struggle when handling specimens with 3D cell groups or dim fluorescent cells. In addition, flow cytometers are often unable to image the cells at all.

Model Theta can capture high resolution (100,000 dpi) 3D images of each single cell with a scanning speed of 24 seconds, and to generate traditional cytometry dot plots and histograms. Both cells in suspension and adherent cells are imaged. In addition, by utilizing in situ sequencing or digital ELISA combined with cytology, important genomic and proteomic parameters can be extracted.


High-Content Screening (HCS)

The ability to identify how novel compounds, peptides or RNAis alter the phenotype of a cell is hugely important in modern drug discovery. High throughput, good resolution and high sensitivity are three critical prerequisites for an efficient HCS system, but are often hard to combine. Although several premium systems nowadays include confocal optics, they lack sensitive cameras. This means in practice that fluorescence imaging is slow with high noise levels and increased photobleaching, especially when taking time-laps images.

Model Theta doesn't compromise when it comes to throughput, sensitivity and resolution, and it's simple to perform the measurements. A plate is imaged at diffraction-limited resolution in 2D in 14.5 minutes with virtually no photobleaching. In addition 3D imaging can easily be performed as well as more complex imaging processing such as spatio-temporal correlation spectroscopy.

We also believe that combining HCS with in situ sequencing or digital ELISA in the way that Model Theta does is a major advance for the drug discovery industry.