Label-free in vivo Raman microspectroscopic imaging of the macromolecular architecture of oocytes

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Confocal Raman spectroscopy (CRS) can provide information about oocyte competency through measurement of changes in the macromolecular architecture during oocyte development and maturation. Hitherto most spectroscopic studies have been limited to fixed oocytes due to the inherent difficulties working with live cells. Here we report the first three-dimensional images of living murine oocytes using CRS. We show that fixation induces significant changes in the macromolecular chemistry compared to living oocytes. A band at 1602 cm-1, assigned to a marker for mitochondria function was found in living oocytes but absent from fixed oocytes providing an in vivo marker. Fixation resulted in significant changes in protein and nucleic acid bands and the spatial distribution of organelles. Raman imaging of Metaphase I and II (MI, MII) and germinal vesicle stage oocytes showed changes in nuclear organisation and cytoplasm macromolecular architecture during these development and maturation stages related to changes in chromosome condensation, mitochondria aggregation and lipid droplet numbers.

Original languageEnglish
Article number8945
Number of pages10
JournalScientific Reports
Volume7
Issue number1
DOIs
Publication statusPublished - 1 Dec 2017

Cite this

@article{c7072181181b4a0bb960f588b9a160bd,
title = "Label-free in vivo Raman microspectroscopic imaging of the macromolecular architecture of oocytes",
abstract = "Confocal Raman spectroscopy (CRS) can provide information about oocyte competency through measurement of changes in the macromolecular architecture during oocyte development and maturation. Hitherto most spectroscopic studies have been limited to fixed oocytes due to the inherent difficulties working with live cells. Here we report the first three-dimensional images of living murine oocytes using CRS. We show that fixation induces significant changes in the macromolecular chemistry compared to living oocytes. A band at 1602 cm-1, assigned to a marker for mitochondria function was found in living oocytes but absent from fixed oocytes providing an in vivo marker. Fixation resulted in significant changes in protein and nucleic acid bands and the spatial distribution of organelles. Raman imaging of Metaphase I and II (MI, MII) and germinal vesicle stage oocytes showed changes in nuclear organisation and cytoplasm macromolecular architecture during these development and maturation stages related to changes in chromosome condensation, mitochondria aggregation and lipid droplet numbers.",
author = "Philip Heraud and Marzec, {Katarzyna Maria} and Qing-Hua Zhang and Yuen, {Wai Shan} and John Carroll and Wood, {Bayden R.}",
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doi = "10.1038/s41598-017-08973-0",
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journal = "Scientific Reports",
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Label-free in vivo Raman microspectroscopic imaging of the macromolecular architecture of oocytes. / Heraud, Philip; Marzec, Katarzyna Maria; Zhang, Qing-Hua; Yuen, Wai Shan; Carroll, John; Wood, Bayden R.

In: Scientific Reports, Vol. 7, No. 1, 8945, 01.12.2017.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Heraud, Philip

AU - Marzec, Katarzyna Maria

AU - Zhang, Qing-Hua

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AU - Carroll, John

AU - Wood, Bayden R.

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AB - Confocal Raman spectroscopy (CRS) can provide information about oocyte competency through measurement of changes in the macromolecular architecture during oocyte development and maturation. Hitherto most spectroscopic studies have been limited to fixed oocytes due to the inherent difficulties working with live cells. Here we report the first three-dimensional images of living murine oocytes using CRS. We show that fixation induces significant changes in the macromolecular chemistry compared to living oocytes. A band at 1602 cm-1, assigned to a marker for mitochondria function was found in living oocytes but absent from fixed oocytes providing an in vivo marker. Fixation resulted in significant changes in protein and nucleic acid bands and the spatial distribution of organelles. Raman imaging of Metaphase I and II (MI, MII) and germinal vesicle stage oocytes showed changes in nuclear organisation and cytoplasm macromolecular architecture during these development and maturation stages related to changes in chromosome condensation, mitochondria aggregation and lipid droplet numbers.

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