Measurement and visualization of cell membrane surface charge in fixed cultured cells related with cell morphology

The diagnosis of patients with malignancies relies on the results of a clinical cytological examination. To enhance the diagnostic qualities of cytological examinations, it is important to have a detailed analysis of the cell’s characteristics. There is, therefore, a need for developing a new auxili...

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Bibliographic Details
Published in:PloS one 2020-07, Vol.15 (7), p.e0236373-e0236373
Main Authors: Nishino, Masaru, Matsuzaki, Ibu, Musangile, Fidele Y., Takahashi, Yuichi, Iwahashi, Yoshifumi, Warigaya, Kenji, Kinoshita, Yuichi, Kojima, Fumiyoshi, Murata, Shin-ichi
Format: Article
Language:English
Subjects:
Amino acids
Anaphase
Antibodies
Apoptosis
Beads
Biology and Life Sciences
Cell adhesion & migration
Cell division
Cell lines
Cell membranes
Cell morphology
Cell surface
Cellular biology
Cytology
Diagnosis
Diagnostic systems
Engineering and Technology
Ion concentration
Ion dynamics
Lipids
Medical diagnosis
Medicine and Health Sciences
Membrane potential
Membranes
Morphology
Pathology
Pepsin
Proteins
Research and Analysis Methods
Supervision
Surface charge
Visualization
Zeta potential
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Summary:The diagnosis of patients with malignancies relies on the results of a clinical cytological examination. To enhance the diagnostic qualities of cytological examinations, it is important to have a detailed analysis of the cell’s characteristics. There is, therefore, a need for developing a new auxiliary method for cytological diagnosis. In this study, we focused on studying the charge of the cell membrane surface of fixed cells, which is one of important cell’s characteristics. Although fixed cells lose membrane potential which is observed in living cells owing to ion dynamics, we hypothesized that fixed cells still have a cell membrane surface charge due to cell membrane components and structure. We used 5 cell lines in this study (ARO, C32TG, RT4, TK, UM-UC-14). After fixation with CytoRich Red, we measured the cell membrane surface charge of fixed cells in solution using zeta potential measurements and fixed cells on glass slides, visualizing it using antibody-labeled beads and positively-charged beads. Furthermore, we measured the cell membrane surface charge of fixed cells under different conditions, such as different solution of fixative, ion concentration, pH, and pepsin treatments. The zeta potential measurements and visualization using the beads indicated that the cell membrane surface of fixed cells was negatively charged, and also that the charge varied among fixed cells. The charge state was affected by the different treatments. Moreover, the number of cell-bound beads was small in interphase, anaphase, and apoptotic cells. We concluded that the negative cell membrane surface charge was influenced by the three-dimensional structure of proteins as well as the different types of amino acids and lipids on the cell membrane. Thus, cell surface charge visualization can be applied as a new auxiliary method for clinical cytological diagnosis. This is the first systematic report of the cell membrane surface charge of fixed cells.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0236373