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Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thaliana
by
Fan, Lusheng
, Hao, Huaiqin
, Wan, Yinglang
, Ash, William M
, Lin, Jinxing
, Kim, Myung K
in
angle of incidence
/ Arabidopsis thaliana
/ Behavior
/ Biological Techniques
/ Biomedical and Life Sciences
/ Boundaries
/ cell wall
/ cell walls
/ cells
/ coat proteins
/ cytosol
/ Fluorescence
/ Fluorescence microscopy
/ Genetic aspects
/ image analysis
/ intact cell
/ Life Sciences
/ Light
/ Methodology
/ Microscopy
/ optical analysis
/ Physiological aspects
/ Plant cells and tissues
/ Plant Sciences
/ Plasma
/ plasma membrane
/ Proteins
/ Quantitative
/ Science
/ Studies
/ Theoretical analysis
/ VA-TIRFM
2011
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Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thaliana
by
Fan, Lusheng
, Hao, Huaiqin
, Wan, Yinglang
, Ash, William M
, Lin, Jinxing
, Kim, Myung K
in
angle of incidence
/ Arabidopsis thaliana
/ Behavior
/ Biological Techniques
/ Biomedical and Life Sciences
/ Boundaries
/ cell wall
/ cell walls
/ cells
/ coat proteins
/ cytosol
/ Fluorescence
/ Fluorescence microscopy
/ Genetic aspects
/ image analysis
/ intact cell
/ Life Sciences
/ Light
/ Methodology
/ Microscopy
/ optical analysis
/ Physiological aspects
/ Plant cells and tissues
/ Plant Sciences
/ Plasma
/ plasma membrane
/ Proteins
/ Quantitative
/ Science
/ Studies
/ Theoretical analysis
/ VA-TIRFM
2011
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While trying to remove the title from your shelf something went wrong :( Kindly try again later!
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Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thaliana
by
Fan, Lusheng
, Hao, Huaiqin
, Wan, Yinglang
, Ash, William M
, Lin, Jinxing
, Kim, Myung K
in
angle of incidence
/ Arabidopsis thaliana
/ Behavior
/ Biological Techniques
/ Biomedical and Life Sciences
/ Boundaries
/ cell wall
/ cell walls
/ cells
/ coat proteins
/ cytosol
/ Fluorescence
/ Fluorescence microscopy
/ Genetic aspects
/ image analysis
/ intact cell
/ Life Sciences
/ Light
/ Methodology
/ Microscopy
/ optical analysis
/ Physiological aspects
/ Plant cells and tissues
/ Plant Sciences
/ Plasma
/ plasma membrane
/ Proteins
/ Quantitative
/ Science
/ Studies
/ Theoretical analysis
/ VA-TIRFM
2011
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Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thaliana
Journal Article
Variable-angle total internal reflection fluorescence microscopy of intact cells of Arabidopsis thaliana
2011
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Overview
Background
Total internal reflection fluorescence microscopy (TIRFM) is a powerful tool for observing fluorescently labeled molecules on the plasma membrane surface of animal cells. However, the utility of TIRFM in plant cell studies has been limited by the fact that plants have cell walls, thick peripheral layers surrounding the plasma membrane. Recently, a new technique known as variable-angle epifluorescence microscopy (VAEM) was developed to circumvent this problem. However, the lack of a detailed analysis of the optical principles underlying VAEM has limited its applications in plant-cell biology.
Results
Here, we present theoretical and experimental evidence supporting the use of variable-angle TIRFM in observations of intact plant cells. We show that when total internal reflection occurs at the cell wall/cytosol interface with an appropriate angle of incidence, an evanescent wave field of constant depth is produced inside the cytosol. Results of experimental TIRFM observations of the dynamic behaviors of phototropin 1 (a membrane receptor protein) and clathrin light chain (a vesicle coat protein) support our theoretical analysis.
Conclusions
These findings demonstrate that variable-angle TIRFM is appropriate for quantitative live imaging of cells in intact tissues of
Arabidopsis thaliana
.
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