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Scan-Free Absorbance Spectral Imaging A(x, y, λ) of Single Live Algal Cells for Quantifying Absorbance of Cell Suspensions.

Isono T, Yamashita K, Momose D, Kobayashi H, Kitamura M, Nishiyama Y, Hosoya T, Kanda H, Kudo A, Okada N, Yagi T, Nakata K, Mineki S, Tokunaga E - PLoS ONE (2015)

Bottom Line: The space-resolved absorbance spectra of the eyespot, an orange organelle about 1 μm, were extracted from the green-color background in a chlorophyll-rich single live cell absorbance image.The formula to calculate the absorbance of cell suspensions from that of single cells was presented to obtain a quantitative, parameter-free agreement with the experiment.It is quantitatively shown that the average number of chlorophylls per cell is significantly underestimated when it is evaluated from the absorbance of the cell suspensions due to the package effect.

View Article: PubMed Central - PubMed

Affiliation: Department of Physics, Faculty of Science, Tokyo University of Science, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162-8601, Japan.

ABSTRACT
Label-free, non-invasive, rapid absorbance spectral imaging A(x,y,λ) microscopy of single live cells at 1.2 μm × 1.2 μm resolution with an NA = 0.85 objective was developed and applied to unicellular green algae Chlamydomonas reinhardtii. By introducing the fiber assembly to rearrange a two-dimensional image to the one-dimensional array to fit the slit of an imaging spectrograph equipped with a CCD detector, scan-free acquisition of three-dimensional information of A(x,y,λ) was realized. The space-resolved absorbance spectra of the eyespot, an orange organelle about 1 μm, were extracted from the green-color background in a chlorophyll-rich single live cell absorbance image. Characteristic absorbance change in the cell suspension after hydrogen photoproduction in C. reinhardtii was investigated to find a single 715-nm absorption peak was locally distributed within single cells. The formula to calculate the absorbance of cell suspensions from that of single cells was presented to obtain a quantitative, parameter-free agreement with the experiment. It is quantitatively shown that the average number of chlorophylls per cell is significantly underestimated when it is evaluated from the absorbance of the cell suspensions due to the package effect.

No MeSH data available.


Related in: MedlinePlus

Experimental data to estimate the spatial resolution of the system.Absorbance at 480 nm around the eyespot in the inset in Fig 4 as a function of position. The FWHM of the absorbance image of the eyespot is about 1.5 μm.
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pone.0128002.g008: Experimental data to estimate the spatial resolution of the system.Absorbance at 480 nm around the eyespot in the inset in Fig 4 as a function of position. The FWHM of the absorbance image of the eyespot is about 1.5 μm.

Mentions: The spatial resolution of the absorbance image is estimated as follows. The spatial distribution of the absorbance image of the eyespot in Fig 4 is displayed in Fig 8.


Scan-Free Absorbance Spectral Imaging A(x, y, λ) of Single Live Algal Cells for Quantifying Absorbance of Cell Suspensions.

Isono T, Yamashita K, Momose D, Kobayashi H, Kitamura M, Nishiyama Y, Hosoya T, Kanda H, Kudo A, Okada N, Yagi T, Nakata K, Mineki S, Tokunaga E - PLoS ONE (2015)

Experimental data to estimate the spatial resolution of the system.Absorbance at 480 nm around the eyespot in the inset in Fig 4 as a function of position. The FWHM of the absorbance image of the eyespot is about 1.5 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465668&req=5

pone.0128002.g008: Experimental data to estimate the spatial resolution of the system.Absorbance at 480 nm around the eyespot in the inset in Fig 4 as a function of position. The FWHM of the absorbance image of the eyespot is about 1.5 μm.
Mentions: The spatial resolution of the absorbance image is estimated as follows. The spatial distribution of the absorbance image of the eyespot in Fig 4 is displayed in Fig 8.

Bottom Line: The space-resolved absorbance spectra of the eyespot, an orange organelle about 1 μm, were extracted from the green-color background in a chlorophyll-rich single live cell absorbance image.The formula to calculate the absorbance of cell suspensions from that of single cells was presented to obtain a quantitative, parameter-free agreement with the experiment.It is quantitatively shown that the average number of chlorophylls per cell is significantly underestimated when it is evaluated from the absorbance of the cell suspensions due to the package effect.

View Article: PubMed Central - PubMed

Affiliation: Department of Physics, Faculty of Science, Tokyo University of Science, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162-8601, Japan.

ABSTRACT
Label-free, non-invasive, rapid absorbance spectral imaging A(x,y,λ) microscopy of single live cells at 1.2 μm × 1.2 μm resolution with an NA = 0.85 objective was developed and applied to unicellular green algae Chlamydomonas reinhardtii. By introducing the fiber assembly to rearrange a two-dimensional image to the one-dimensional array to fit the slit of an imaging spectrograph equipped with a CCD detector, scan-free acquisition of three-dimensional information of A(x,y,λ) was realized. The space-resolved absorbance spectra of the eyespot, an orange organelle about 1 μm, were extracted from the green-color background in a chlorophyll-rich single live cell absorbance image. Characteristic absorbance change in the cell suspension after hydrogen photoproduction in C. reinhardtii was investigated to find a single 715-nm absorption peak was locally distributed within single cells. The formula to calculate the absorbance of cell suspensions from that of single cells was presented to obtain a quantitative, parameter-free agreement with the experiment. It is quantitatively shown that the average number of chlorophylls per cell is significantly underestimated when it is evaluated from the absorbance of the cell suspensions due to the package effect.

No MeSH data available.


Related in: MedlinePlus