Fe-TAMLs as a new class of small molecule peroxidase probes for correlated light and electron microscopy.
Stephen R AdamsMason R MackeyRanjan RamachandraThomas J DeerinckGuillaume A CastillonSebastien PhanJunru HuDaniela BoassaJohn T NgoMark H EllismanPublished in: bioRxiv : the preprint server for biology (2023)
We introduce Fe-TAML, a small molecule-based peroxidase as a versatile new member of the correlated fluorescence and electron microscopy toolkit. The utility of the probe is demonstrated by high resolution imaging of newly synthesized DNA (through biorthogonal labeling), genetically tagged proteins (using HaloTag), and untagged endogenous proteins (via immunostaining). EM visualization in these applications is facilitated by exploiting Fe-TAML's catalytic activity for the deposition of localized osmiophilic precipitates based on polymerized 3,3'-diaminobenzidine. Optimized conditions for synthesizing and implementing Fe-TAML based probes are also described. Overall, Fe-TAML is a new chemical biology tool that can be used to visualize diverse biomolecular species along nanometer and micron scales within cells.
Keyphrases
- small molecule
- electron microscopy
- high resolution
- single molecule
- protein protein
- metal organic framework
- living cells
- aqueous solution
- hydrogen peroxide
- induced apoptosis
- visible light
- mass spectrometry
- nucleic acid
- nitric oxide
- quality improvement
- quantum dots
- endoplasmic reticulum stress
- cell free
- cell cycle arrest
- pi k akt