Figure 2: Cells are labeled with CMOs in a stepwise process.

First, the cholesterol-modified Universal Anchor Strand is pre-hybridized with the Adapter Strand. Next, the Universal Anchor + Adapter solution is mixed with the cell suspension. The cholesterol on the Universal Anchor + Adapter complex inserts into the cell membrane. After incubation, the cholesterol-modified Universal Co-Anchor Strand is added to the cell suspension, where it hybridizes with the Universal Anchor Strand and inserts into the cell membrane. The addition of the second cholesterol molecule increases the net hydrophobicity of the DNA complex and stabilizes it within the membrane²⁶. After washing out the excess DNA, the cells are concentrated and added to a PDMS flow cell on top of the patterned surface. The 3’ end of the Adapter Strand hybridizes with the Surface DNA Strand on the glass slide, resulting in adhesion to the slide specifically in regions functionalized with complementary DNA.