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Updated: Jun 8, 2026

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy
Published on: March 16, 2020
Covalent immobilization of protein onto a functionalized hydrogenated diamond-like carbon substrate
Hari Shankar Biswas1, Jagannath Datta, D P Chowdhury
1Nanocrystalline Diamond like Carbon Synthesis Laboratory, Saha Institute of Nuclear Physics, 1/AF, Bidhan Nagar, Kolkata-700 064, India.
Abstract:
Hydrogenated diamond-like carbon (HDLC) has an atomically smooth surface that can be deposited on high-surface area substrata and functionalized with reactive chemical groups, providing an ideal substrate for protein immobilization. A synthetic sequence is described involving deposition and hydrogenation of DLC followed by chemical functionalization. These functional groups are reacted with amines on proteins causing covalent immobilization on contact. Raman measurements confirm the presence of these surface functional groups, and Fourier transform infrared spectroscopy (FTIR) confirms covalent protein immobilization. Atomic force microscopy (AFM) of immobilized proteins is reproducible because proteins do not move as a result of interactions with the AFM probe-tip, thus providing an advantage over mica substrata typically used in AFM studies of protein. HDLC offers many of the same technical advantages as oxidized graphene but also allows for coating large surface areas of biomaterials relevant to the fabrication of medical/biosensor devices.

