Multicolor multiphoton in vivo imaging flow cytometry.

Lingjie Kong,Jianyong Tang,Meng Cui

doi:10.1364/oe.24.006126

Abstract

In vivo flow cytometry provides a non-invasive way of probing the biology of circulating cells during disease progression and studying cellular response to therapy. However, current methods provide little morphological information which potentially could be new biological marker for early disease diagnosis, and fail to reveal intercellular interactions. Here we report a multi-color, multiphoton in vivo imaging flow cytometry, to image circulating cells within the vasculature of scattering tissues at high spatiotemporal resolution. We apply it in imaging of cellular dynamics in bone marrow through the intact mouse skull, in situ deformability cytometry, distinguishing cellular clusters, and simultaneously monitoring multiple types of trafficking cells based on their morphologies and fluorescence emission colors.

Highlights

Traditional flow cytometry (FC) is used routinely to acquire quantitative information about specific cell populations at high throughput, and has found extensive clinical applications in diagnostic pathology [1, 2]
After going through the polarization beam splitter (PBS), quarter waveplate (QWP) and ultrasound lenses (UL, TAG lens 2.0, TAG Optics), the beams were imaged onto a mirror (M1) by a pair of relay lens (RL1 and RL1’)
The reflected beams from M1 went through the UL and QWP again, and were reflected by the PBS as their polarizations were rotated by 90° after the second pass of the QWP

Summary

Introduction

Traditional flow cytometry (FC) is used routinely to acquire quantitative information about specific cell populations at high throughput, and has found extensive clinical applications in diagnostic pathology [1, 2]. Detection and quantification of rare circulating cells in vivo are very important for early diagnosis of diseases (such as cancer, stroke, and inflammation), and long-term study is necessary for probing the biology of circulating cells during disease progression, and for studying cellular response to therapy (such as drugs and radiation) [1, 3,4,5,6,7,8,9] To this end, several methods of in vivo FC have been developed to continuously monitor circulating cells in live animals without affecting the physiology of the subject, and found applications in many biomedical studies, such as cancer, immunology, and stem cells [3,4,5,10,11,12]. A drawback of in vivo FC is that it generally provides little morphological information (such as the size, shape, morphology, and deformability of circulating cells), which can potentially be new biological marker that is sensitive to early disease development [13, 14]

Methods

Results

Conclusion