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<p>Electron cryomicroscopy (cryo-EM) allows high spatial resolution visualization of biological specimens; however, it is challenging to chemically identify densities observed in cryo-EM. To overcome this, we combined cryo-EM with chemical imaging using focused ion beam secondary ion mass spectrometry (FIB-SIMS) for integrated spatiochemical analysis of untagged specimens. We show that our correlative workflow permits subcellular localization of molecules inside bacterial cells and is compatible with cryogenic light microscopy and FIB-milled lamellae of eukaryotic specimens. To highlight biological insights enabled by the workflow, we studied the uptake of bisphenol-AF, a widespread chemical pollutant, by environmental bacteria, revealing the storage of these chemicals within cytosolic phase-separated aggregates in pollutant-exposed cells, where they cannot be removed by the bacterial efflux machinery despite its robust upregulation. Cryo-EM-FIB-SIMS therefore represents an effective approach to map elemental and molecular signatures in near-native biological samples.</p>

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Subcellular chemical mapping using correlated cryogenic electron and mass spectrometry imaging

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月25日

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  • Hannah Ochner,
  • Buse Isbilir,
  • Sonja Blasche,
  • David Scheidweiler,
  • Yuexuan Zhang,
  • Zhexin Wang,
  • Tom Smith,
  • Catarina Franco,
  • Rob Bradley,
  • Kiran R. Patil,
  • Tanmay A. M. Bharat

摘要

Electron cryomicroscopy (cryo-EM) allows high spatial resolution visualization of biological specimens; however, it is challenging to chemically identify densities observed in cryo-EM. To overcome this, we combined cryo-EM with chemical imaging using focused ion beam secondary ion mass spectrometry (FIB-SIMS) for integrated spatiochemical analysis of untagged specimens. We show that our correlative workflow permits subcellular localization of molecules inside bacterial cells and is compatible with cryogenic light microscopy and FIB-milled lamellae of eukaryotic specimens. To highlight biological insights enabled by the workflow, we studied the uptake of bisphenol-AF, a widespread chemical pollutant, by environmental bacteria, revealing the storage of these chemicals within cytosolic phase-separated aggregates in pollutant-exposed cells, where they cannot be removed by the bacterial efflux machinery despite its robust upregulation. Cryo-EM-FIB-SIMS therefore represents an effective approach to map elemental and molecular signatures in near-native biological samples.