New: Our latest case study on qFibrosis' role in the successful development of Rezdiffra

SHG Microscopy for Collagen Imaging

Stain-Free Precision

SHG microscopy is a laser-based imaging technique that allows visualization of non-centrosymmetric structures like fibrillar collagen in tissues without the use of exogenous stains or dyes. This optical phenomenon occurs when two photons interact with a non-centrosymmetric structure and combine to generate a new photon with exactly twice the energy and frequency.

Revealing Hidden Structures - How SHG works

Next-generation Imaging

In SHG microscopy, tissues are exposed to femtosecond laser pulses. When these pulses interact with fibrillar collagen, the asymmetry in the structure causes the combining of two lower energy photons into a single higher energy photon. This emitted photon has exactly half the wavelength of the excitation laser. Special filters enable detecting these emitted photons to generate high contrast, high resolution images showing the distribution of fibrillar collagen.


As SHG signal generation is a physics phenomenon rather than one that depends on chemical dyes or stains, it avoids issues with staining variation, photobleaching and provides output images with intrinsic contrast in a highly consistent manner. The SHG intensity is also directly proportional to the quantity and organization of fibrillar collagen, allowing quantitative analysis.

Enabling Non-Destructive Collagen Monitoring

Continuous Fibrosis Profiling

Compared to traditional histological stains like Picosirius Red or Masson’s Trichrome, SHG provides intrinsic signals specific to fibrillar collagen structure. This eliminates variability due to stain penetration or protocol differences.

Since there is no photobleaching, repeated imaging is possible without signal loss. Furthermore, the non-destructive nature also preserves tissues for subsequent analyses. Quantification of SHG signals provides objective measures of fibrillar collagen content and organization without observer interpretation biases of staining intensity.

SHG Imaging MT Stain

Extensive Applications Across Diseases

Fibrotic Diseases

  • Liver Fibrosis
  • Renal Fibrosis
  • Pulmonary Fibrosis
  • Bone Marrow Fibrosis
  • Dermal Fibrosis
  • Sub-epithelial Fibrosis on Cornea
  • Retinal Pigment Epithelium
  • Arterial Fibrosis
  • Cardiac Fibrosis
  • Biliary Disease
  • Muscular Dystrophy (Sarcopenia)
  • Alcoholic Liver Disease
  • Alpha-1 antitrypsin Deficiency 
  • Biliary Atresia


  • Breast Cancer
  • Pancreatic Cancer
  • Liver Cancer
  • Lung Cancer
  • Ovarian Cancer
  • Colon Cancer
  • Kidney Cancer
  • Bladder Cancer
  • Squamous Cell Carcinoma of the Head and Neck (SCCHN)

Ushering in a New Era of Tissue Evaluation

Transforming Imaging in Pathology

Supplementing SHG microscopy with AI assessments, our platforms analyze liver biopsies to accurately quantify fibrosis – unlocking continuous metrics that sensitively detect subtle histological changes that may be missed by stains. The continuous values have shown good correlation with conventional pathology and objectively enhanced disease characterization, reduce variability in assessments, and provide greater sensitivity to accurately measure the magnitude of treatment efficacy.