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( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a <t>living</t> <t>COS-7</t> cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.
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( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a <t>living</t> <t>COS-7</t> cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.
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( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a <t>living</t> <t>COS-7</t> cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.
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( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a <t>living</t> <t>COS-7</t> cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.
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( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a living COS-7 cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.

Journal: Science Advances

Article Title: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)

doi: 10.1126/sciadv.aed9613

Figure Lengend Snippet: ( A ) Schematic of the dynaMIET experiment conducted on a PM-labeled cell on top of a MIET substrate. ( B and E ) Same as , but for the PM system. n = 40 independent measurements over three independent experiments for living cells, and n = 25 independent measurements over two independent experiments for fixed cells. Data are obtained from independent measurements on multiple cells. For each cell, only one to two independent measurements were collected to avoid pseudo-replication. Independent experiments refer to measurements performed on different days using independently prepared samples. ( C and F ) Box plots of mean height values h 0 , diffusion coefficients D , fluctuation amplitudes ψ , and relaxation times τ ∗ for living and fixed cells. ( D ) Reconstructed 3D topography of the fluorescently labeled basal PM of a living COS-7 cell on a MIET substrate. The bottom panel offers an alternative viewing angle. Height is color-coded for clarity.

Article Snippet: COS-7 monkey kidney cell (American Type Culture Collection, CRL-1651) was a gift from S. Rizzoli (University Medicine Göttingen).

Techniques: Labeling, Diffusion-based Assay

( A , B , C , E , and F ) Same as but for the ER system. n = 25 independent measurements over 3 independent experiments for both living cells and fixed cells. Experimental replication and statistical details are the same as described in . ( D ) Exemplary 3D reconstructions (height maps) of fluorescently labeled ER in a live COS-7 cell on a MIET substrate. The xy image shows the fluorescence intensity. The bottom panel shows the height image from a different viewing angle.

Journal: Science Advances

Article Title: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)

doi: 10.1126/sciadv.aed9613

Figure Lengend Snippet: ( A , B , C , E , and F ) Same as but for the ER system. n = 25 independent measurements over 3 independent experiments for both living cells and fixed cells. Experimental replication and statistical details are the same as described in . ( D ) Exemplary 3D reconstructions (height maps) of fluorescently labeled ER in a live COS-7 cell on a MIET substrate. The xy image shows the fluorescence intensity. The bottom panel shows the height image from a different viewing angle.

Article Snippet: COS-7 monkey kidney cell (American Type Culture Collection, CRL-1651) was a gift from S. Rizzoli (University Medicine Göttingen).

Techniques: Labeling, Fluorescence

( A to F ) Same as but for the NE system. n = 33 independent measurements over three independent experiments. Experimental replication and statistical details are the same as described in . (B) iACFs obtained from living eGFP-nup153–modified COS-7 cell. The green solid line represents a fit with . The inset shows measured g i for a fixed cell. (E) Calculated hACF for a living and a fixed cell (inset).

Journal: Science Advances

Article Title: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)

doi: 10.1126/sciadv.aed9613

Figure Lengend Snippet: ( A to F ) Same as but for the NE system. n = 33 independent measurements over three independent experiments. Experimental replication and statistical details are the same as described in . (B) iACFs obtained from living eGFP-nup153–modified COS-7 cell. The green solid line represents a fit with . The inset shows measured g i for a fixed cell. (E) Calculated hACF for a living and a fixed cell (inset).

Article Snippet: COS-7 monkey kidney cell (American Type Culture Collection, CRL-1651) was a gift from S. Rizzoli (University Medicine Göttingen).

Techniques: Modification