H-EM: An algorithm for simultaneous cell diameter and intensity quantification in low-resolution imaging cytometry.

Area of Science:

• Biomedical Engineering
• Cell Biology
• Image Analysis

Background:

• Flow cytometry and microscopy are standard cell analysis techniques.
• Low-magnification "cell astronomy" offers simplicity and high throughput but faces signal-to-noise challenges.
• Previous cell astronomy systems struggled with quantitative cell-labeling measurements.

Purpose of the Study:

• To develop a cell astronomy system capable of reliable cell intensity and diameter quantification.
• To introduce a novel algorithm (H-EM) for accurate cell analysis at low magnification.
• To demonstrate the system's efficacy with calibrated beads and stained cells.

Main Methods:

• Development of a low-magnification "cell astronomy" imaging system.
• Custom algorithm development, including Fisher discriminant analysis and a novel H-EM algorithm.
• Validation using calibrated MESF beads and fluorescently stained leukocytes.

Main Results:

• The cell astronomy system reliably quantifies cell intensities and diameters.
• The H-EM algorithm achieves accurate brightness and diameter estimates with sub-pixel precision.
• Good population identification was achieved for both beads and leukocytes, even in clusters.

Conclusions:

• The developed cell astronomy system and H-EM algorithm overcome limitations of low-magnification imaging.
• This approach enables accurate and high-throughput cell analysis, advancing image-based cytometry.
• The H-EM algorithm offers improved performance over existing methods like DAOSTORM for cell quantification.