⭐ High Impact

A multimodal platform for nonlinear optical microscopy and microspectroscopy.

Chen Hongtao, Wang Haifeng, Slipchenko Mikhail N, Jung YooKyung, Shi Yunzhou, Zhu Jiabin, Buhman Kimberly K, Cheng Ji-Xin

📰 Optics express 📅 2009 📊 85 citations
DOI PubMed

Abstract

Multimodal nonlinear optical microscopy is a valuable tool to study complex biological samples. We present an easy-to-operate approach to perform coherent anti-Stokes Raman scattering (CARS), two-photon fluorescence (TPF), second harmonic generation (SHG), and third-harmonic generation (THG) imaging using a single laser source composed of an 80 MHz femtosecond (fs) laser, an optical parametric oscillator (OPO), and a PPLN crystal for frequency doubling. The platform allows vibrationally resonant CARS imaging of CH-rich myelin sheath in fresh spinal tissues and lipid bodies in live cells. Multimodal nonlinear optical imaging and microspectroscopy analysis of fresh liver tissues are demonstrated.

🔬 Techniques

Coherent Anti-Stokes Raman Scattering Confocal Laser Scanning Microscopy Live Cell Imaging Second Harmonic Generation Z-Stack Imaging

🔭 Microscopes

FV1000 Olympus FV1000 Confocal

💻 Software

FluoView

🧪 Sample Preparation

Live Imaging

🏭 Microscope Brands

Coherent Olympus Hamamatsu Chroma Spectra-Physics

🧪 Reagent Suppliers

MatTek Sigma-Aldrich

🔴 Lasers

Spectra-Physics Mai Tai Ti:Sapphire

📷 Detectors

Hamamatsu PMT PMT

🎨 Filters

430/40 bandpass 520/40 bandpass 650/45 bandpass

💻 Software Details

Image Acquisition:
FluoView
Image Analysis:
ImageJ

🏛️ Research Organizations (ROR)

Affiliated research institutions:

🏛 Purdue University West Lafayette

📋 Methods

✔ Verified methods section 2,592 words Read on PMC ↗

Sample preparation TiO 2 nanopowder (

📊 Figures

Fig. 1

Fig. 1

Schematic diagram of a multimodal NLO microscope using a fs laser source. The Mai Tai output at 790 nm (u03c9 1 ) is split into two beams, with 80% of power being used to pump the Opal-BB. The idler o...

Fig. 2

Fig. 2

Spatial resolution. (a) Epi-detected CARS image of TiO 2 nanoparicles, excited at 790 and 1018 nm. (b-c) Lateral and axial intensity profiles along the circled TiO 2 nanoparticle in (a). (d) Epi-detec...

Fig. 3

Fig. 3

Vibrational contrast of C-H rich objects. (a-d) The forward-detected CARS images of a subcutaneous fat tissue by the fs (a-b) and ps (c-d) lasers, respectively. The Raman shift is marked in each image...

Fig. 4

Fig. 4

Comparison of forward-detected THG (a) and CARS (b) images of lipid droplets inside live KB cells. Scale bars = 10 u03bcm.

Fig. 5

Fig. 5

Multimodal nonlinear optical imaging of fresh liver tissues from high fat fed mice. (a) TPF image excited at 790 nm and detected in the 450-550 nm region. Fibrous structure on the liver surface produc...

Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.

🏛️ Imaging Facility

🏛️ Purdue University

💬 Discussion

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