使用Fiji

DAB Hematoxylin分離

Image->Colour->Colour Deocnvolution

選擇"H DAB"

計算細胞數

使用Hematoxylin圖

Threshold->Auto->Apply

Analyze->Analyze Particles，set "Size=100-Infinity"✓ "Summarize"

定量DAB

Type->8bit

Invert+Threshold (Dark background)+Set measurement (show Area&Mean gray value, check "limited to threshold")

"Measure"以獲得Mean&Area

算法1: Intensity/nuclei

(Mean*Area)/nuclei#

算法2: Intensity/Area

References

1. https://www.researchgate.net/post/Does_anyone_have_a_protocol_for_quantifying_IHC_images_in_ImageJ