Label-Free Imaging of Single Proteins Secreted from Living Cells via iSCAT Microscopy

Journal article


Publication Details

Author(s): Gemeinhardt A, McDonald MP, König K, Aigner M, Mackensen A, Sandoghdar V
Journal: Journal of Visualized Experiments
Publication year: 2018
Journal issue: 141
ISSN: 1940-087X


Abstract

We demonstrate interferometric scattering (iSCAT) microscopy, a method capable of detecting single unlabeled proteins secreted from individual living cells in real time. In this protocol, we cover the fundamental steps to realize an iSCAT microscope and complement it with additional imaging channels to monitor the viability of a cell under study. Following this, we use the method for real-time detection of single proteins as they are secreted from a living cell which we demonstrate with an immortalized B-cell line (Laz388). Necessary steps concerning the preparation of microscope and sample as well as the analysis of the recorded data are discussed. The video protocol demonstrates that iSCAT microscopy offers a straightforward method to study secretion at the single-molecule level.


FAU Authors / FAU Editors

Mackensen, Andreas Prof. Dr.
Lehrstuhl für Innere Medizin V


External institutions with authors

Helmholtz-Zentrum Berlin für Materialien und Energie (HZB)


How to cite

APA:
Gemeinhardt, A., McDonald, M.P., König, K., Aigner, M., Mackensen, A., & Sandoghdar, V. (2018). Label-Free Imaging of Single Proteins Secreted from Living Cells via iSCAT Microscopy. Journal of Visualized Experiments, 141. https://dx.doi.org/10.3791/58486

MLA:
Gemeinhardt, André, et al. "Label-Free Imaging of Single Proteins Secreted from Living Cells via iSCAT Microscopy." Journal of Visualized Experiments 141 (2018).

BibTeX: 

Last updated on 2019-22-07 at 07:16