Characteristics of genetic tags for correlative light and electron microscopy

Beatty, Kimberly E.; López, Clàudia

doi:10.1016/j.cbpa.2023.102369

Cited by 4 publications

(1 citation statement)

References 52 publications

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“…Probes and tags for visualizing biomolecules by light and electron microscopy are powerful tools for dissecting the molecular organization of cell [1,2]. Over recent years, molecular-engineering based strategies been used to develop new labels to track specific proteins via EM, including small taggable peptide sequences or proteins [3][4][5][6], fluorescent proteins capable of generating EM-visible reaction products [7] as well as enzyme-based tags such as encodable peroxidases HRP and APEX2 [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Fe-TAMLs as a new class of small molecule peroxidase probes for correlated light and electron microscopy

Adams,

Mackey,

Ramachandra

et al. 2023

Preprint

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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.

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