Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy

@article{Denk1994AnatomicalAF,
  title={Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy},
  author={W. Denk and K. Delaney and A. Gelperin and D. Kleinfeld and B. Strowbridge and D. Tank and R. Yuste},
  journal={Journal of Neuroscience Methods},
  year={1994},
  volume={54},
  pages={151-162}
}
Light scattering by brain tissue and phototoxicity are major obstacles to the use of high-resolution optical imaging and photo-activation ('uncaging') of bioactive compounds from inactive ('caged') precursors in intact and semi-intact nervous systems. Optical methods based on 2-photon excitation promise to reduce these obstacles (Denk, 1994; Denk et al., 1990, 1994). Here we show a range of imaging modes based on 2-photon laser scanning microscopy (TPLSM) as applicable to problems in… Expand
Supraresolution Imaging in Brain Slices using Stimulated-Emission Depletion Two-Photon Laser Scanning Microscopy
TLDR
STED 2PLSM supraresolution microscopy achieves approximately 3-fold improvement in resolution in the radial direction over conventional 2PL SM, revealing greater detail in the structure of dendritic spines located approximately 100 microns below the surface of brain slices. Expand
Cortical imaging through the intact mouse skull using two‐photon excitation laser scanning microscopy
TLDR
In an effort to image the brain with subcellular spatial resolution and without the “cortical window” procedure, a method was devised to image directly through thinned mouse skull using two-photon excitation microscopy. Expand
Multiphoton Imaging of Neurons in Living Tissue: Acquisition and Analysis of Time-Lapse Morphological Data
TLDR
Object-Image, an extension of the popular public-domain NIH Image software, together with a set of custom macros for time-lapse morphometric analysis, is used to analyze neuronal branch growth and complexity in three dimensions over time. Expand
Optical Imaging of Neural Structure and Physiology: Confocal Fluorescence Microscopy in Live Brain Slices
TLDR
Multiphoton imaging provides intrinsic 3D resolution while confining fluorescence excitation to a single narrow focal plane, thus reducing the risk of photodynamic damage. Expand
Chapter 10. In vivo measurements of blood flow and glial cell function with two-photon laser-scanning microscopy.
TLDR
The dual role of two-photon imaging as a means to assess function in the normal state as well as a tool to investigate the vascular system and glia under pathological conditions, such as ischemia and microvascular disease are emphasized. Expand
Two-photon microscopy in brain tissue: parameters influencing the imaging depth
TLDR
An improved low-magnification, high-numerical aperture objective is proposed that should allow fluorescence measurements related to neuronal or vascular brain activity at >100 microm deeper than with standard objectives. Expand
Two-photon microscopy : sequential imaging studies in vivo
Microscopists have always desired to look inside various organ tissues to study structure, function and dysfunction of their cellular constituents. In the past, this has frequently required tissueExpand
Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain
TLDR
In Scale-treated mouse brain, neurons labeled with genetically encoded fluorescent proteins were visualized at an unprecedented depth in millimeter-scale networks and at subcellular resolution, suggesting that the Scale method will be useful for light microscopy–based connectomics of cellular networks in brain and other tissues. Expand
Principles of Two-Photon Excitation Microscopy and Its Applications to Neuroscience
TLDR
The principles of 2PE microscopy are reviewed, recent applications are highlighted, its limitations are discussed, and areas for future research and development are pointed to. Expand
Two-photon excitation in functional biological imaging.
  • W. Denk
  • Physics, Medicine
  • Journal of biomedical optics
  • 1996
TLDR
The contribution that the combination of two-photon absorption with laser scanning20 has made to functional biological imaging in recent years is reviewed. Expand
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 39 REFERENCES
Infrared-interference videomicroscopy of living brain slices.
  • H. Dodt
  • Materials Science, Medicine
  • Advances in experimental medicine and biology
  • 1993
TLDR
The novel combination of infrared and DIC described here permits visualization of neurons in tissue slices of 300–500 μm conventionally employed for in vitro electrophysiological analysis and is referred to in the following as interference optics. Expand
Photostimulation using caged glutamate reveals functional circuitry in living brain slices.
  • E. Callaway, L. Katz
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1993
TLDR
Caged glutamate-based photostimulation eliminates artifacts and limitations inherent in conventional stimulation methods, including stimulation of axons of passage, desensitization, and poor temporal resolution of "puffer" pipettes, and current artifacts of iontophoretic application. Expand
Two-photon scanning photochemical microscopy: mapping ligand-gated ion channel distributions.
  • W. Denk
  • Chemistry, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1994
TLDR
It is shown that the highly localized liberation of "caged" neurotransmitters by two-photon absorption-mediated photoactivation can be used in conjunction with recording the induced whole-cell current to determine the distribution of ligand-gated ion channels. Expand
Direct observation of neurotoxicity in brain slices with infrared videomicroscopy
TLDR
By recording in time lapse mode it was possible to visualize the dynamics of cell swelling and to demonstrate neuroprotection by glutamatergic antagonists and the method may be of use in screening of potential neuroprotective drugs for stroke therapy. Expand
Two-photon laser scanning fluorescence microscopy.
TLDR
The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation. Expand
Cortical point-spread function and long-range lateral interactions revealed by real-time optical imaging of macaque monkey primary visual cortex
TLDR
The usefulness of real-time optical imaging in the study of population activity and the exploration of cortical dendritic processing is described, raising the possibility that distributed processing over a very large cortical area plays a major role in the processing of visual information by the primary visual cortex of the primate. Expand
Optical mapping of electrical activity in rat somatosensory and visual cortex
TLDR
Simultaneous, multi-site optical recordings of activity may provide a new and potentially powerful method for studying function and dysfunction in mammalian cortex. Expand
Mapping calcium transients in the dendrites of Purkinje cells from the guinea‐pig cerebellum in vitro.
TLDR
The results indicate that the absorbance signals came from calcium entry into the cell resulting from the turning on of voltage‐dependent calcium conductances, and suggests that calcium spikes causing these signals can be evoked separately in different regions of the Purkinje cell dendritic field by long‐lasting potentials which may reach local threshold at different times. Expand
Identification of presynaptic neurons by laser photostimulation.
TLDR
An optical method involving the use of a laser and a novel fluorescent dye as a photostimulation probe has been developed to identify presynaptic neurons in a large ensemble of cells, speeding up the elucidation of neuronal networks. Expand
NONLINEAR ABSORPTION EXTENDS CONFOCAL FLUORESCENCE MICROSCOPY INTO THE ULTRA-VIOLET REGIME AND CONFINES THE ILLUMINATION VOLUME
It is shown that two-photon absorption confines the illumination volume and present quantitative evidence that an additional confocal arrangement of the detector further improves the resolution byExpand
...
1
2
3
4
...