Two-photon imaging of spatially extended neuronal network dynamics with high temporal resolution

Journal of Neuroscience Methods
Kyle P LillisJerome Mertz


We describe a simple two-photon fluorescence imaging strategy, called targeted path scanning (TPS), to monitor the dynamics of spatially extended neuronal networks with high spatiotemporal resolution. Our strategy combines the advantages of mirror-based scanning, minimized dead time, ease of implementation, and compatibility with high-resolution low-magnification objectives. To demonstrate the performance of TPS, we monitor the calcium dynamics distributed across an entire juvenile rat hippocampus (>1.5mm), at scan rates of 100 Hz, with single cell resolution and single action potential sensitivity. Our strategy for fast, efficient two-photon microscopy over spatially extended regions provides a particularly attractive solution for monitoring neuronal population activity in thick tissue, without sacrificing the signal-to-noise ratio or high spatial resolution associated with standard two-photon microscopy. Finally, we provide the code to make our technique generally available.


May 1, 1988·Epilepsy Research·T J Chesnut, J W Swann
Dec 14, 1999·Journal of Neuroscience Methods·Y P TanE Neher
Sep 27, 2002·Biophysical Journal·James D LechleiterIlse Sieneart
Jun 5, 2003·Proceedings of the National Academy of Sciences of the United States of America·Christoph StosiekArthur Konnerth
Nov 5, 2003·Nature Biotechnology·Warren R ZipfelWatt W Webb
Mar 5, 2004·Journal of Neurophysiology·Robert D RoordaGero Miesenböck
Nov 22, 2005·Nature Methods·Fritjof Helmchen, Winfried Denk
Jan 31, 2006·Journal of Neuroscience Methods·Rafael KurtzMarkus Sauer
Feb 7, 2006·Journal of Neuroscience Methods·R SaloméL Bourdieu
Dec 5, 2006·Nature Methods·Werner GöbelFritjof Helmchen
Apr 5, 2007·Nature Protocols·Olga GaraschukArthur Konnerth
Oct 30, 2007·Nature Methods·Volodymyr NikolenkoRafael Yuste
Dec 19, 2007·Optics Letters·J BewersdorfS W Hell
Mar 8, 2008·Applied Optics·K H KimP T So
Jun 12, 2006·Optics Express·Ran YiJerome Mertz


Aug 11, 2012·Nature·Nathan R WilsonMriganka Sur
May 25, 2013·Nature Protocols·Nathan R WilsonMriganka Sur
Jun 27, 2013·Proceedings of the National Academy of Sciences of the United States of America·Mathieu DucrosSerge Charpak
Jul 7, 2010·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·David Parker
Jul 1, 2011·Journal of Neurophysiology·Alexander J SadovskyJason N MacLean
Jul 9, 2010·Journal of Neurophysiology·Ilya ValmianskiDavid Kleinfeld
Mar 22, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Matt WachowiakMarkus Rothermel
Mar 2, 2012·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Helen R SabolekKevin J Staley
May 9, 2012·The Review of Scientific Instruments·Runhua JiangShaoqun Zeng
May 1, 2009·The Journal of Physiology·F HaissB Weber
Mar 5, 2013·Optics Letters·Jorge Pérez-VizcaínoJesús Lancis
May 12, 2009·Journal of Neuroscience Methods·Karolina Szokol, Marie-Claude Perreault
Feb 2, 2012·Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism·Andy Y ShihDavid Kleinfeld
Mar 18, 2015·Epilepsia·Kyle P LillisJokūbas Žiburkus
Jun 9, 2012·Neurobiology of Disease·Kyle P LillisJohn A White
Oct 27, 2009·Current Opinion in Neurobiology·Benjamin F Grewe, Fritjof Helmchen
Dec 3, 2014·Journal of Neuroscience Methods·Mohammad I K HamadPetra Wahle
Dec 17, 2014·Journal of Neuroscience Methods·Serena Bovetti, Tommaso Fellin
Jul 12, 2014·Progress in Neurobiology·Stefano FumagalliMaria-Grazia De Simoni
Aug 30, 2016·Nature Neuroscience·Na JiSpencer L Smith
Jan 6, 2017·Scientific Reports·Serena BovettiTommaso Fellin
Nov 14, 2017·Journal of Neural Engineering·Renaud SchuckSimon R Schultz
Aug 3, 2013·Cold Spring Harbor Protocols·Jonathan D DriscollDavid Kleinfeld
Dec 18, 2020·Frontiers in Neuroscience·Meng-Jung Lee, Günther Zeck
Jan 13, 2021·Proceedings of the National Academy of Sciences of the United States of America·Yayao MaLiang Gao

Related Concepts

Structure of Subiculum Hippocampi
Resting Potentials
Neural Networks (Anatomic)
Neural Pathways
Organ Culture Techniques
Computer Programs and Programming

Trending Feeds


Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Genetic Screens in iPSC-derived Brain Cells

Genetic screening is a critical tool that can be employed to define and understand gene function and interaction. This feed focuses on genetic screens conducted using induced pluripotent stem cell (iPSC)-derived brain cells. It also follows CRISPR-Cas9 approaches to generating genetic mutants as a means of understanding the effect of genetics on phenotype.

Alzheimer's Disease: MS4A

Variants within the membrane-spanning 4-domains subfamily A (MS4A) gene cluster have recently been implicated in Alzheimer's disease in genome-wide association studies. Here is the latest research on Alzheimer's disease and MS4A.

Pediculosis pubis

Pediculosis pubis is a disease caused by a parasitic insect known as Pthirus pubis, which infests human pubic hair, as well as other areas with hair including eye lashes. Here is the latest research.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Rh Isoimmunization

Rh isoimmunization is a potentially preventable condition that occasionally is associated with significant perinatal morbidity or mortality. Discover the latest research on Rh Isoimmunization here.

Pharmacology of Proteinopathies

This feed focuses on the pharmacology of proteinopathies - diseases in which proteins abnormally aggregate (i.e. Alzheimer’s, Parkinson’s, etc.). Discover the latest research in this field with this feed.

Enzyme Evolution

This feed focuses on molecular models of enzyme evolution and new approaches (such as adaptive laboratory evolution) to metabolic engineering of microorganisms. Here is the latest research.

Alignment-free Sequence Analysis Tools

Alignment-free sequence analyses have been applied to problems ranging from whole-genome phylogeny to the classification of protein families, identification of horizontally transferred genes, and detection of recombined sequences. Here is the latest research.