Assessment of Population and ECM Production Using Multiphoton Microscopy as an Indicator of Cell Viability

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1601)

Abstract

Multiphoton microscopy allows continuous depth-resolved, nondestructive imaging of scaffold-seeded cells during cell or tissue culture. Spectrally separated images in high resolution can be provided while cells are conserved in their native state. Here we describe the seeding of mesenchymal stem cells to bacterial nanocellulose hydropolymer scaffolds followed by 2-channel imaging of cellular autofluorescence (AF) and collagen-I formation using second harmonic generation (SHG) signals. With this approach the simultaneous observation of the progression of cell morphology and production of extracellular matrix as hallmarks of viability and cell fitness is possible.

Key words

Multiphoton imaging Cell viability Extracellular matrix formation Collagen-I SHG Cellular autofluorescence Scaffold Bacterial nanocellulose 

Notes

Acknowledgments

This work was supported by the Emerging Fields Initiative (EFI) of the University of Erlangen-Nürnberg (project TOPbiomat) and the Erlangen Graduate School in Advanced Optical Technologies (SAOT) within the German Excellence Initiative. We want to thank Dana Kralisch and Nadine Hessler (JeNaCell GmbH) for providing BNC fleeces.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Friedrich-Alexander University (FAU) Erlangen-NürnbergInstitute of Medical BiotechnologyErlangenGermany

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