Skip to main content
Journal cover image

Evaluation of a method to measure fluorescent cell burden in complex culture systems.

Publication ,  Journal Article
Holden, R; Park, J; Price, A; Floyd, S; Oldham, M
Published in: Biomed Phys Eng Express
April 27, 2022

Purpose. This work introduces and evaluates a method for accurate in-vitro measurement of fluorescent cell burden in complex 3D-culture conditions.Methods.The Fluorescent Cell Burden (FCB) method was developed to analyze the burden of 4T1 mCherry-expressing cells grown in an organotypic co-culture model of brain metastasis using 400μm rat brain slices. As a first step, representative simulated image-data accurately reflecting the 4T1 experimental data, but with known ground truth burden, were created. The FCB method was then developed in the CellProfiler software to measure the integrated intensity and area of the colonies in the simulated image data. Parameters in the pipeline were varied to span the experimentally observed range (e.g. of cell colony size) and the result compared with simulation ground truth to evaluate and optimize FCB performance. The optimized CellProfiler pipeline was then applied to the original 4T1 tumor cell images to determine colony growth with time, and re-applied with upper and lower bound parameters to determine uncertainty estimates.Results.The FCB method measured integrated intensity across 10 simulated images with an accuracy of 99.23% ± 0.75%. When colony density was increased by increasing colony number to 450, 600, and 750, the FCB measurement was 98.68%, 100.9%, 97.6% and 113.5% of the true value respectively. For the increasing number of cells plated on the rat brain slices, the integrated intensity increased nearly linearly with cell count except for at high cell counts, where it is hypothesized that shadowing from clumped cells causes a sub-linear relationship.Conclusion. The FCB method accurately measured an integrated fluorescent light intensity to within 5% of ground truth for a wide range of simulated image data spanning the range of observed variability in experimental data. The method is readily customizable to in-vitro studies requiring estimation of fluorescent tumor cell burden.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Biomed Phys Eng Express

DOI

EISSN

2057-1976

Publication Date

April 27, 2022

Volume

8

Issue

3

Location

England

Related Subject Headings

  • Software
  • Humans
  • Computer Simulation
  • Brain Neoplasms
  • 4003 Biomedical engineering
  • 3206 Medical biotechnology
  • 1004 Medical Biotechnology
  • 0903 Biomedical Engineering
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Holden, R., Park, J., Price, A., Floyd, S., & Oldham, M. (2022). Evaluation of a method to measure fluorescent cell burden in complex culture systems. Biomed Phys Eng Express, 8(3). https://doi.org/10.1088/2057-1976/ac6701
Holden, R., J. Park, A. Price, S. Floyd, and M. Oldham. “Evaluation of a method to measure fluorescent cell burden in complex culture systems.Biomed Phys Eng Express 8, no. 3 (April 27, 2022). https://doi.org/10.1088/2057-1976/ac6701.
Holden R, Park J, Price A, Floyd S, Oldham M. Evaluation of a method to measure fluorescent cell burden in complex culture systems. Biomed Phys Eng Express. 2022 Apr 27;8(3).
Holden, R., et al. “Evaluation of a method to measure fluorescent cell burden in complex culture systems.Biomed Phys Eng Express, vol. 8, no. 3, Apr. 2022. Pubmed, doi:10.1088/2057-1976/ac6701.
Holden R, Park J, Price A, Floyd S, Oldham M. Evaluation of a method to measure fluorescent cell burden in complex culture systems. Biomed Phys Eng Express. 2022 Apr 27;8(3).
Journal cover image

Published In

Biomed Phys Eng Express

DOI

EISSN

2057-1976

Publication Date

April 27, 2022

Volume

8

Issue

3

Location

England

Related Subject Headings

  • Software
  • Humans
  • Computer Simulation
  • Brain Neoplasms
  • 4003 Biomedical engineering
  • 3206 Medical biotechnology
  • 1004 Medical Biotechnology
  • 0903 Biomedical Engineering