Last but not least: this achievement was only possible due to the amazing dedication and insight by @dijo-mj.bsky.social, @linussison.bsky.social, @nazlicanyurekli.bsky.social, and Sheston Culpepper!

This methodology adds a fundamentally new perspective to our understanding of cell biology. We’re looking forward to ideas and feedback – please get in touch!

Have you ever wondered where different sialylation states of the EGF receptor are located on the cell surface? Here you go:

This enables us to map glycoforms of individual proteins as well as their organization on the cell membrane in the native state.

Here, we demonstrate the first spatial mapping of the cell-surface glycoproteome at true molecular resolution. We target proteins of interest with antibody-nanobody constructs and address glycosylation with either lectins or metabolic oligosaccharide engineering.

Various methods like mass spectrometry have been used to study the cell-surface glycoproteome with great success, however, no technique could analyze the spatial organization of proteins and their glycosylation patterns at molecular resolution.

Every cell in the human body is surrounded by the glycocalyx, the "sugar coat" of the cell. A key component of the glycocalyx are glycosylated proteins. Indeed, virtually all cell-surface proteins are glycosylated.

Maybe I’m biased: but well deserved:) 🙌

Geeking out is the biggest compliment:D

Huge shoutout to @dijo-mj.bsky.social and @nazlicanyurekli.bsky.social who spearheaded this study! Also, thanks to our amazing collaborators Sarah Fritsche, Reem Hashem, Oana-Maria Thoma, Imen Larafa, Tina Boric, Chloe Bielawski, @karimalmahayni.bsky.social, Kristian Franze, and Maximilan Waldner!

Glycan Atlassing links glycocalyx structure to cell function, and we are looking forward to applying this strategy to exciting questions in fundamental and clinical glycoscience!

Strikingly, we found that glycan patterns communicate cell state via the glycocalyx. We can make these visible: Below, each dot is one cell, and each color is a different stage of cancer progression. Just looking at the glycocalyx, we can see which cell is at which stage in the oncogenic cascade.

These images look pretty, but can we do more with them? Turns out, we can! We developed an analysis pipeline to understand how the different glycan species talk to each other. It turns out: They are organized in highly specific ways on the cell surface.

With this, we obtained an “atlas of glycans” on a range of sample types, from cultured cells over primary immune cells to neurons and patient tissue. Each dot in the image below is one glycan (sub-)unit, resolved at the nanometer scale.

We tackled this problem by labeling different glycan units within the glycocalyx using lectins. These lectins were tagged with DNA barcodes, which we used for multiplexed DNA-PAINT super-resolution microscopy.