Breast Cancer ECM Visualization using Multiphoton Imaging

Jeremy BredfeldtLOCI, Morgridge Institute for

Research

H&E and SHG

• Protocol– Thaw & fix sample– Section sample to 200 microns on a vibratome– Image with SHG– Paraffin embed tissue wafer– Section and stain for H&E– Slide scan

• Green = SHG signal (Collagen Type I)

Normal Associated Tissue

11.6 mm

H&E

SHG

Invasive Ductal Carcinoma

H&E

SHG

Fibroadenoma

H&E SHG

7.5 mm

Mucinous Carcinoma

H&ESHG

Intrinsic Fluorescence

• Green = SHG• Red = Autofluorescence from FAD or Collagen

type III

FibroAdenoma

Extrinsic Fluorescence

• Green = SHG (Collagen type I)• Red = E-Cadherin stained cells &

Autofluorescence from Collagen type III

Terminal Ductal Lobular Unit (Red) Surrounded by Collagen (Green)

Duct in Cross Section, Surrounded by IDC cells

IDC

Duct

IDC Cells Near a Vessel with ECM

IDC Cells

Vessel