Quantification of cell surface receptor expression in live tissue culture media using a dual-tracer stain and rinse approach

Xiaochun Xu; Lagnojita Sinha; Aparna Singh; Cynthia Yang; Jialing Xiang; Kenneth M. Tichauer

doi:10.1117/12.2078472

2 March 2015 Quantification of cell surface receptor expression in live tissue culture media using a dual-tracer stain and rinse approach

Xiaochun Xu, Lagnojita Sinha, Aparna Singh, Cynthia Yang, Jialing Xiang, Kenneth M. Tichauer

Author Affiliations +

Proceedings Volume 9328, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIII; 932814 (2015) https://doi.org/10.1117/12.2078472
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

Immunofluorescence staining is a robust way to visualize the distribution of targeted biomolecules invasively in in fixed tissues and tissue culture. Despite the fact that these methods has been a well-established method in fixed tissue imaging for over 70 years, quantification of receptor concentration still simply assumes that the signal from the targeted fluorescent marker after incubation and sufficient rinsing is directly proportional to the concentration of targeted biomolecules, thus neglecting the experimental inconsistencies in incubation and rinsing procedures and assuming no, nonspecific binding of the fluorescent markers. This work presents the first imaging approach capable of quantifying the concentration of cell surface receptor on cancer cells grown in vitro based on compartment modeling in a nondestructive way. The approach utilizes a dual-tracer protocol where any non-specific retention or variability in incubation and rinsing of a receptor-targeted imaging agent is corrected by simultaneously imaging the retention of a chemically similar, “untargeted” imaging agent. Various different compartment models were used to analyze the data in order to find the optimal procedure for extracting estimates of epidermal growth factor receptor (EGFR) concentration (a receptor overexpressed in many cancers and a key target for emerging molecular therapies) in tissue cultures with varying concentrations of human glioma cells (U251). Preliminary results demonstrated a need to model nonspecific binding of both the targeted and untargeted imaging agents used. The approach could be used to carry out the first repeated measures of cell surface receptor dynamics during 3D tumor mass development, in addition to the receptor response to therapies.

Citation Download Citation

Xiaochun Xu, Lagnojita Sinha, Aparna Singh, Cynthia Yang, Jialing Xiang, and Kenneth M. Tichauer "Quantification of cell surface receptor expression in live tissue culture media using a dual-tracer stain and rinse approach", Proc. SPIE 9328, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIII, 932814 (2 March 2015); https://doi.org/10.1117/12.2078472

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

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
6 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Receptors

Tissues

Data modeling

Natural surfaces

Tumor growth modeling

3D modeling

Cancer

Show All Keywords

Keywords/Phrases

Search In:

Publication Years