Detail of Fig3E_HEK293T_PBS



Project
Title
Superresolution structured illumination microscopy (SR-SIM) images of HEK293T cells expressing membrane EGFP stained with DAPI (nuclei) and MitoTracker (mitochondria) in PBS samples
Description
NA
Release, Updated
2017-10-03,
2018-11-15
License
CC BY
Kind
Image data based on Experiment
File Formats
Data size
764.9 MB

Organism
H. sapiens
Strain(s)
HEK293T
Cell Line
-
Protein names
membrane, nucleus, mitochondria
Protein tags
EGFP
Reporter
DAPI, MitoTracker

Datatype
neuron structure
Molecular Function (MF)
Biological Process (BP)
-
Cellular Component (CC)
-
Biological Imaging Method
XYZ Scale
XY: 0.025 micrometer/pixel, Z: 0.109 micrometer/slice
T scale
-

Image Acquisition
Experiment type
Other
Microscope type
ConfocalMicroscope
Acquisition mode
LaserScanningConfocalMicroscopy
Contrast method
Fluorescence
Microscope model
Carl Zeiss ELYRA PS.1 (SR-SIM)
Detector model
EM-CCD
Objective model
Carl Zeiss alpha Plan-Apochromat 100x/1.46 O
Filter set
-

Summary of Methods
See details in Ke et al. (2016) Cell Reports, 14: 2718-2732.
Related paper(s)

Meng-Tsen Ke, Yasuhiro Nakai, Satoshi Fujimoto, Rie Takayama, Shuhei Yoshida, Tomoya S Kitajima, Makoto Sato, Takeshi Imai (2016) Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent., Cell reports, Volume 14, Number 11, pp. 2718-32

Published in 2016 Mar 22 (Electronic publication in March 10, 2016, midnight )

(Abstract) Super-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we report an optimized optical clearing agent for high-resolution fluorescence imaging (SeeDB2). SeeDB2 matches the refractive indices of fixed tissues to that of immersion oil (1.518), thus minimizing both light scattering and spherical aberrations. During the clearing process, fine morphology and fluorescent proteins were highly preserved. SeeDB2 enabled super-resolution microscopy of various tissue samples up to a depth of >100 mum, an order of magnitude deeper than previously possible under standard mounting conditions. Using this approach, we demonstrate accumulation of inhibitory synapses on spine heads in NMDA-receptor-deficient neurons. In the fly medulla, we found unexpected heterogeneity in axon bouton orientations among Mi1 neurons, a part of the motion detection circuitry. Thus, volumetric super-resolution microscopy of cleared tissues is a powerful strategy in connectomic studies at synaptic levels.
(MeSH Terms)

Contact
Takeshi Imai , RIKEN , Center for Developmental Biology , Laboratory for Sensory Circuit Formation
Contributors
Meng-Tsen Ke, Yasuhiro Nakai, Satoshi Fujimoto, Rie Takayama, Shuhei Yoshida, Tomoya S. Kitajima, Makoto Sato, Takeshi Imai

OMERO Dataset
OMERO Project
Source