Photoemission electron microscopy for connectomics

G Gregg Wildenberg (Department of Neurobiology, University of Chicago) K Kevin M. Boergens (Department of Physics, University of Illinois Chicago) L Lola Lambert (Department of Neurobiology, University of Chicago) R Ruiyu Li (Department of Chemistry, University of Chicago) A Allison Craig (James Franck Institute, University of Chicago) M Michael K. L. Man (Okinawa Institute of Science and Technology, Femtosecond Spectroscopy Unit) A Amin Moradi (Leiden Institute of Physics, Leiden University) J Janek Rieger (Department of Chemistry, University of Chicago) H Hengli Duan (Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, United Kingdom) S Sarnjeet S. Dhesi (Diamond Light Source, Magnetic Materials Group) G Gabriel Karras F Francesco Maccherozzi (Diamond Light Source, Magnetic Materials Group) K Keshav Dani (Okinawa Institute of Science and Technology, Femtosecond Spectroscopy Unit) R Rudolf Tromp (Leiden Institute of Physics, Leiden University) S Sense Jan van der Molen (Leiden Institute of Physics, Leiden University) S Sarah B. King (Department of Chemistry) N Narayanan Kasthuri (Department of Neurobiology, University of Chicago)

Abstract

Photoemission electron microscopy (PEEM) offers a potential third modality for large-volume connectomics alongside transmission electron microscopy (TEM) and scanning electron microscopy (SEM). We image osmium stained, ultrathin brain sections on gold coated silicon at synaptic resolution using commercial PEEMs. At coarser resolution, we demonstrate that ultraviolet laser illumination enables gigavoxel-per-second acquisition rates without thermal damage. PEEM combines TEM-like parallel detection with SEM-compatible solid supports into a potentially scalable and cost-effective approach for large-volume connectomes.

Article Details

Volume / Issue Vol. 122, Issue 48
Published December 02, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (17)

G

Gregg Wildenberg

Department of Neurobiology, University of Chicago

K

Kevin M. Boergens

Department of Physics, University of Illinois Chicago

L

Lola Lambert

Department of Neurobiology, University of Chicago

R

Ruiyu Li

Department of Chemistry, University of Chicago

A

Allison Craig

James Franck Institute, University of Chicago

M

Michael K. L. Man

Okinawa Institute of Science and Technology, Femtosecond Spectroscopy Unit

A

Amin Moradi

Leiden Institute of Physics, Leiden University

J

Janek Rieger

Department of Chemistry, University of Chicago

H

Hengli Duan

Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, United Kingdom

S

Sarnjeet S. Dhesi

Diamond Light Source, Magnetic Materials Group

G

Gabriel Karras

F

Francesco Maccherozzi

Diamond Light Source, Magnetic Materials Group

K

Keshav Dani

Okinawa Institute of Science and Technology, Femtosecond Spectroscopy Unit

R

Rudolf Tromp

Leiden Institute of Physics, Leiden University

S

Sense Jan van der Molen

Leiden Institute of Physics, Leiden University

S

Sarah B. King

Department of Chemistry

N

Narayanan Kasthuri

Department of Neurobiology, University of Chicago