Quantitative Nanoscale Secondary Ion Mass Spectrometry (NanoSIMS) Imaging of Individual Vesicles to Investigate the Relation between Fraction of Chemical Release and Vesicle Size

• Published in: Angewandte Chemie Vol. 62; no. 28; pp. e202304098 - n/a
• Main Authors: Nguyen, Tho Duc Khanh, Rabasco, Stefania, Lork, Alicia A., Toit, Andre Du, Ewing, Andrew G.
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 10.07.2023
• Edition: International ed. in English
• Subjects: Cell Membrane; Chemical Release Fraction; Chemical Sciences; Diagnostic Imaging; Dopamine; Exocytosis; Ions; Kemi; Mass spectrometry; Mass spectroscopy; Nanosims; Partial Release; Scientific imaging; Secondary ion mass spectrometry; Spectrometry, Mass, Secondary Ion; TEM; Transmission electron microscopy; Vesicles; Chemical Release Fraction; Partial Release; Exocytosis; TEM; Nanosims
• ISSN: 1433-7851; 1521-3773; 0044-8249; 1521-3757; 1521-3773
• DOI: 10.1002/anie.202304098

We used correlative transmission electron microscopy (TEM) and nanoscale secondary ion mass spectrometry (NanoSIMS) imaging to quantify the contents of subvesicular compartments, and to measure the partial release fraction of 13C‐dopamine in cellular nanovesicles as a function of size. Three modes of exocytosis comprise full release, kiss‐and‐run, and partial release. The latter has been subject to scientific debate, despite a growing amount of supporting literature. We tailored culturing procedures to alter vesicle size and definitively show no size correlation with the fraction of partial release. In NanoSIMS images, vesicle content was indicated by the presence of isotopic dopamine, while vesicles which underwent partial release were identified by the presence of an 127I‐labelled drug, to which they were exposed during exocytosis allowing entry into the open vesicle prior to its closing again. Demonstration of similar partial release fractions indicates that this mode of exocytosis is predominant across a wide range of vesicle sizes. Correlative transmission electron microscopy (TEM) and nanoscale secondary ion mass spectrometry (NanoSIMS) imaging were used to measure vesicle size and vesicular isotopic dopamine content in cellular vesicles. Measurements before and after stimulated exocytotic release show the fraction of chemical release is independent of vesicle size.