Fluorescence Dynamics in the Endoplasmic Reticulum of a Live Cell: Time‐Resolved Confocal Microscopy |
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Authors: | Dr. Shirsendu Ghosh Somen Nandi Catherine Ghosh Prof. Dr. Kankan Bhattacharyya |
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Affiliation: | Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata, India), Fax: (91)-33-2473-2805 |
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Abstract: | Fluorescence dynamics in the endoplasmic reticulum (ER) of a live non‐cancer lung cell (WI38) and a lung cancer cell (A549) are studied by using time‐resolved confocal microscopy. To selectively study the organelle, ER, we have used an ER‐Tracker dye. From the emission maximum of the ER‐Tracker dye, polarity (i.e. dielectric constant, ?) in the ER region of the cells (≈500 nm in WI38 and ≈510 nm in A549) is estimated to be similar to that of chloroform ( =506 nm, ?≈5). The red shift by 10 nm in in the cancer cell (A549) suggests a slightly higher polarity compared to the non‐cancer cell (WI38). The fluorescence intensity of the ER‐Tracker dye exhibits prolonged intermittent oscillations on a timescale of 2–6 seconds for the cancer cell (A549). For the non‐cancer cell (WI38), such fluorescence oscillations are much less prominent. The marked fluorescence intensity oscillations in the cancer cell are attributed to enhanced calcium oscillations. The average solvent relaxation time (<τs>) of the ER region in the lung cancer cell (A549, 250±50 ps) is about four times faster than that in the non‐cancer cell (WI38, 1000±50 ps). |
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Keywords: | confocal microscopy endoplasmic reticulum fluorescence oscillation lung cells solvent relaxation |
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