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448-Temma-HeLaSPASIM/Fig2_onephoton_mito_fixed_SPASIM thumbnail

448-Temma-HeLaSPASIM/Fig2_onephoton_mito_fixed_SPASIM

Author reviewed
Fluorescence images of mitochondria in fixed HeLa cells in the imaging mode of SPA-SIM for single-photon activation
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Project
448-Temma-HeLaSPASIM
Dataset ID
Fig2_onephoton_mito_fixed_SPASIM
Kind
Image data
Title
Fluorescence images of mitochondria in fixed HeLa cells in the imaging mode of SPA-SIM for single-photon activation
Description
The fluorescence images of mitochondria labeled with Skylan-NS in fixed HeLa cells were taken and reconstructed in the imaging mode of selective-plane-activation structured illumination microscopy (SPA-SIM). The SPA-SIM mode produced large improvements in image contrast and spatial resolution, even for a densely labeled thick specimen. SPA-SIM reduced the influence of the out-of-focus background more effectively, which facilitates the visualization of intracellular structures. Skylan-NS is a photoswitchable green fluorescent protein. CW laser is used for single-photon activation, which is more efficient and allows measurement with higher temporal resolution.
Folder Name
Fig2_onephoton_mito_fixed_SPASIM
File Format
tif
File Size
9MB
File Separation Method
-
Figure Number
Figure2d
Dimensions
3069x3069x1x1x1
X Scale
70.9 micrometer
Y Scale
70.9 micrometer
Z Scale
-
T Scale
-
Channel
1 channel
Quantitative Data
-

Biosample

Description
Mitochondria were visualized using mitochondria-targeted Skylan-NS in living HeLa cell spheroids.
Organism
Homo sapiens
(NCBITaxon:9606)
Strain
-
Cell
-
Cell line
HeLa-S3 cell (CLO:0003699)
MeSH
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Mitochondria (D008928)
Intrinsic Variable
-
Extrinsic Variable
HeLa cells were electroporated with Skylan-NS plasmids

Ontology

Anatomical Entity
-
UBERON
-
Biological Process
-
Cellular Component
mitochondrion (GO:0005739)
Molecular Function
-
MeSH Anatomy
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Mitochondria (D008928)
MeSH BP
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Mitochondria (D008928)
MeSH CC
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Mitochondria (D008928)
MeSH MF
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Mitochondria (D008928)

Genetics

Gene
-
Protein / Probe
-
Method / Genotype
electroporation

Treatments

Reagent / Compound
-
Concentration
-
Fold Dilution
-

Imaging Method

Imaging Method
structured illumination microscopy
(FBbi:00000332)
Detection Method
detection of photons
Visualization Method
Skylan-NS
Illumination Method
light-sheet illumination
Source of Contrast
-
Imaged Parameter
-
Sample Preparation Method
fixed cell
MeSH
HeLa Cells (D006367)
Microscopy, Fluorescence (D008856)
Mitochondria (D008928)

Instrument

Body
-
Module
-
Light Source
405 nm light sheet illumination
Detector
Hamamatsu Photonics ORCA-Flash 4.0 v3
Objective
Nikon CFI75 Apo 25XC W, Special Optics 54-10-7
Filter Set
-
Dichroic
Di03-R488-t3-25x36, RazorEdge LongPass488