Improved forward scatter detection of a flow cytometer for detection of extracellular vesicles

P. R. Bloemen; L. de Rond; E. van der Pol; T. G. van Leeuwen; F. A. W. Coumans

doi:10.1117/12.2527919

19 July 2019 Improved forward scatter detection of a flow cytometer for detection of extracellular vesicles

P. R. Bloemen, L. de Rond, E. van der Pol, T. G. van Leeuwen, F. A. W. Coumans

Proceedings Volume 11073, Clinical and Preclinical Optical Diagnostics II; 1107313 (2019) https://doi.org/10.1117/12.2527919
Event: European Conferences on Biomedical Optics, 2019, Munich, Germany

Abstract

Extracellular vesicles (EVs) are nanoparticles secreted from cells into bodily fluids. EVs are potential biomarkers for diseases such as thrombosis or cancer. However, the small size and low refractive index of EVs complicates their detection. A flow cytometer is suited for EV characterisation, but typically lacks scatter sensitivity on one or both scatter detectors for derivation of both particle size and refractive index. Here, we aim to improve the FACSCanto (Becton Dickinson) forward scatter detector for the detection of 100 nm EVs, which requires an improvement in SNR of 10⁷-fold based on Mie theory. This was achieved through replacement of the 20 mW laser by an 200 mW laser, replacement of the photodiode detector with a photomultiplier tube and a confocalized optical geometry. Using a prototype optical setup, we obtained an improvement in SNR which was 1,11·10⁴ – fold better than the standard design. However, the optics was suboptimal and far from diffraction-limited. Zemax simulations led to a nearly diffraction limited optical design which is expected to yield another 200-fold improvement. Taken together these changes will improve the SNR 2.2·10⁶-fold and thus improve the detection limit of the FACSCanto to 130 nm EVs.

Citation Download Citation

P. R. Bloemen, L. de Rond, E. van der Pol, T. G. van Leeuwen, and F. A. W. Coumans "Improved forward scatter detection of a flow cytometer for detection of extracellular vesicles", Proc. SPIE 11073, Clinical and Preclinical Optical Diagnostics II, 1107313 (19 July 2019); https://doi.org/10.1117/12.2527919

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available