Core calculations
All calculations are derived from Digman et al. (2008).
Here img
represents a numpy array of shape (t, y, x)
.
Intensity
Intensity is calculated as:
In python:
average_intensity = np.mean(img, axis=0)
Variance
Variance is calculated as:
In python:
variance = np.var(img, axis=0)
Apparent brightness
Apparent brightness is calculated as:
In python:
apparent_brightness = variance / average_intensity
Apparent number
Apparent number is calculated as:
In python:
apparent_number = average_intensity**2 / variance
Brightness
Brightness is calculated as:
In python:
brightness = (variance - average_intensity) / (average_intensity - background)
Number
Number is calculated as:
In python:
number = ((average_intensity-background)**2) / np.clip((variance - average_intensity), 1e-6, None)
Here the denominator is clipped (limited) to a value of 1e-6 to prevent extremely high number values.
References:
- Digman, M. A., Dalal, R., Horwitz, A. F., & Gratton, E. (2008). Mapping the Number of Molecules and Brightness in the Laser Scanning Microscope. Biophysical Journal, 94(6), 2320–2332. https://doi.org/10.1529/biophysj.107.114645