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Fluorescence Dynamics in the Endoplasmic Reticulum of a Live Cell: Time‐Resolved Confocal Microscopy
Authors:Dr. Shirsendu Ghosh  Somen Nandi  Catherine Ghosh  Prof. Dr. Kankan Bhattacharyya
Affiliation:Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata, India), Fax: (91)-33-2473-2805
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 urn:x-wiley:14394235:media:cphc201600425:cphc201600425-math-0001 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 (urn:x-wiley:14394235:media:cphc201600425:cphc201600425-math-0002 =506 nm, ?≈5). The red shift by 10 nm in urn:x-wiley:14394235:media:cphc201600425:cphc201600425-math-0003 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).
Keywords:confocal microscopy  endoplasmic reticulum  fluorescence oscillation  lung cells  solvent relaxation
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