Alexa 594 labelled GFAP primary adult human astrocytes

A novel assay using a combination of morphology filters, thresholding and image morphometry analysis (IMA) to quantify cells that have intriguingly complex morphology.

• Especially suited to primary cells, like astrocytes stained for GFAP, or microglia stained for CD45 (see below).
• Quantification of GFAP positive primary adult human astrocytes.

Original image.		Thresholded image.		Cells quantified using IMA and standard object size configuration (this divides clumped or overlapping cells based on specified average cell size).

Alexa 488 labelled Nestin primary adult human astrocytes

Quantification of nestin positive primary adult human astrocytes.

Original image.		Morphology filter applied to image to improve signal to noise ratio.		Positive cell marked in green. Orange objects failed to meet the area and staining intensity filter range, these objects were therefore excluded as noise and background staining.

DAB labelled CD45 primary adult human microglia

Using primary adult human microglia stained for CD45 with DAB instead of fluorescence.

Original image (captured with transmitted light).		Invert objects morphology filters applied to convert pixels to fluorescence scale for better contrast and to remove noise.		Border objects morphology filters applied to convert pixels to fluorescence scale for better contrast and to remove noise.
Image thresholded and IMA applied

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