Histochemistry is a microscopy-based technology widely used to visualize the molecular distribution in biological tissue. Recent developments in label-free optical imaging has demonstrated the potential to replace the conventional histochemical labels/markers (fluorescent antibodies, organic dyes, nucleic acid probes, and other contrast agents) with diverse optical interactions to generate histochemical contrasts, allowing “virtual” histochemistry in three spatial dimensions without preparing a microscope slide (i.e. labor-intensive sample preparation). However, the histochemical information in a label-free optical image has often been rather limited due to the difficulty in simultaneously generating multiple histochemical contrasts with strict spatial co-registration. Here, in the first part (Part I) of this two-part series study, we develop a technique of slide-free virtual histochemistry based on label-free multimodal multiphoton microscopy, and simultaneously generate up to four histochemical contrasts from in vivo animal and ex vivo human tissue. To enable this functionality, we construct and demonstrate a robust fiber-based laser source for clinical translation and phenotype a wide variety of vital cells in unperturbed mammary tissue.

Original languageEnglish (US)
Article number#332416
Pages (from-to)5240-5252
Number of pages13
JournalBiomedical Optics Express
Issue number11
StatePublished - Nov 1 2018

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics


Dive into the research topics of 'Slide-free virtual histochemistry (Part I): Development via nonlinear optics'. Together they form a unique fingerprint.

Cite this