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Single-shot full-field reflection phase microscopy.

Yaqoob Z, Yamauchi T, Choi W, Fu D, Dasari RR, Feld MS - Opt Express (2011)

Bottom Line: The reflection-based DHM provides highly sensitive and a single-shot imaging of cellular dynamics while the use of low coherence source provides a depth-selective measurement.The setup uniquely uses a diffraction grating in the reference arm to generate an interference image of uniform contrast over the entire field-of-view albeit low-coherence light source.We have measured the path-length sensitivity of our instrument to be approximately 21 picometers/Hz that makes it suitable for nanometer-scale full-field measurement of membrane dynamics in live cells.

View Article: PubMed Central - PubMed

Affiliation: G. R. Harrison Spectroscopy Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. zyaqoob@mit.edu

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Setup and results of the cell membrane fluctuation measurement. (a) Location of coherence gate; the sample is tilted to simultaneously acquire membrane fluctuations as well as background phase from the coverslip. (b) Power spectral density of membrane fluctuations as a function of frequency for three different populations: blue, formalin fixed; green, normal; and red, Cytochalasin-D treated HeLa cells.
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g005: Setup and results of the cell membrane fluctuation measurement. (a) Location of coherence gate; the sample is tilted to simultaneously acquire membrane fluctuations as well as background phase from the coverslip. (b) Power spectral density of membrane fluctuations as a function of frequency for three different populations: blue, formalin fixed; green, normal; and red, Cytochalasin-D treated HeLa cells.

Mentions: To demonstrate our system’s capabilities, we have studied the membrane fluctuations in HeLa cells under different cell conditions. More specifically, we prepared (i) a sample of living normal HeLa cells, (ii) a fixed HeLa cell sample after treatment with 2% paraformaldehyde and (iii) a sample of HeLa cells treated with 8 nM Cytochalasin-D which inhibits actin polymerization [25]. The frame rate of the image acquisition was set to 1 kHz and the data was recorded for duration of 1 sec for each cell. As shown in Fig. 5(a)Fig. 5


Single-shot full-field reflection phase microscopy.

Yaqoob Z, Yamauchi T, Choi W, Fu D, Dasari RR, Feld MS - Opt Express (2011)

Setup and results of the cell membrane fluctuation measurement. (a) Location of coherence gate; the sample is tilted to simultaneously acquire membrane fluctuations as well as background phase from the coverslip. (b) Power spectral density of membrane fluctuations as a function of frequency for three different populations: blue, formalin fixed; green, normal; and red, Cytochalasin-D treated HeLa cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3368324&req=5

g005: Setup and results of the cell membrane fluctuation measurement. (a) Location of coherence gate; the sample is tilted to simultaneously acquire membrane fluctuations as well as background phase from the coverslip. (b) Power spectral density of membrane fluctuations as a function of frequency for three different populations: blue, formalin fixed; green, normal; and red, Cytochalasin-D treated HeLa cells.
Mentions: To demonstrate our system’s capabilities, we have studied the membrane fluctuations in HeLa cells under different cell conditions. More specifically, we prepared (i) a sample of living normal HeLa cells, (ii) a fixed HeLa cell sample after treatment with 2% paraformaldehyde and (iii) a sample of HeLa cells treated with 8 nM Cytochalasin-D which inhibits actin polymerization [25]. The frame rate of the image acquisition was set to 1 kHz and the data was recorded for duration of 1 sec for each cell. As shown in Fig. 5(a)Fig. 5

Bottom Line: The reflection-based DHM provides highly sensitive and a single-shot imaging of cellular dynamics while the use of low coherence source provides a depth-selective measurement.The setup uniquely uses a diffraction grating in the reference arm to generate an interference image of uniform contrast over the entire field-of-view albeit low-coherence light source.We have measured the path-length sensitivity of our instrument to be approximately 21 picometers/Hz that makes it suitable for nanometer-scale full-field measurement of membrane dynamics in live cells.

View Article: PubMed Central - PubMed

Affiliation: G. R. Harrison Spectroscopy Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. zyaqoob@mit.edu

Show MeSH
Related in: MedlinePlus