Issue 12, 2023

Saline dry fixation for improved cell composition analysis using Raman spectroscopy

Abstract

Raman spectroscopy enables the label-free assessment of cellular composition. While live cell analysis is the most accurate approach for cellular Raman spectroscopy, the analysis of fixed cells has proved to be very useful, particularly in collaborative projects where samples need to be serially examined by different laboratories or stored and reanalyzed at a later date. However, many chemicals that are widely used for cell fixation directly affect cellular biomolecules, yielding Raman spectra with missing or altered information. In this article, we compared the suitability of dry-fixation with saline versus chemical fixatives. We compared the Raman spectroscopy of saline dry-fixed cells with the more commonly used formaldehyde and methanol fixation and found that dry-fixed cell spectra preserved more cellular information than either chemical fixative. We also assessed the stability of dry-fixed cells over time and found that they were stable for at least 5 months. Finally, a comparison of dry-fixed and live cell spectra revealed effects due to the hydration state of the cells since they were recovered upon rehydrating dry-fixed samples. Thus, for fixed cell Raman spectroscopy, we recommend dry-fixation with unbuffered saline as a superior method to formaldehyde or methanol fixation.

Graphical abstract: Saline dry fixation for improved cell composition analysis using Raman spectroscopy

Supplementary files

Article information

Article type
Paper
Submitted
29 ربيع الثاني 1444
Accepted
21 شوال 1444
First published
24 شوال 1444

Analyst, 2023,148, 2745-2757

Saline dry fixation for improved cell composition analysis using Raman spectroscopy

S. Rangan, R. Wong, H. G. Schulze, M. Z. Vardaki, M. W. Blades, R. F. B. Turner and J. M. Piret, Analyst, 2023, 148, 2745 DOI: 10.1039/D2AN01916G

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements