In Vivo Simultaneous Nonlinear Absorption Raman and Fluorescence (SNARF) Imaging of Mouse Brain Cortical Structures

Overview

Journal Commun Biol

Specialty Biology

Date 2022 Mar 11

PMID 35273325

Authors

Andrew T Francis

Bryce Manifold

Elena C Carlson

Ruoqian Hu

Andrew H Hill

Shuaiqian Men

Dan Fu

Affiliations

Soon will be listed here.

Abstract

Label-free multiphoton microscopy is a powerful platform for biomedical imaging. Recent advancements have demonstrated the capabilities of transient absorption microscopy (TAM) for label-free quantification of hemoglobin and stimulated Raman scattering (SRS) microscopy for pathological assessment of label-free virtual histochemical staining. We propose the combination of TAM and SRS with two-photon excited fluorescence (TPEF) to characterize, quantify, and compare hemodynamics, vessel structure, cell density, and cell identity in vivo between age groups. In this study, we construct a simultaneous nonlinear absorption, Raman, and fluorescence (SNARF) microscope with the highest reported in vivo imaging depth for SRS and TAM at 250-280 μm to enable these multimodal measurements. Using machine learning, we predict capillary-lining cell identities with 90% accuracy based on nuclear morphology and capillary relationship. The microscope and methodology outlined herein provides an exciting route to study several research topics, including neurovascular coupling, blood-brain barrier, and neurodegenerative diseases.

Citing Articles

Quantitative Optical Imaging of Oxygen in Brain Vasculature.

Rathbone E, Fu D J Phys Chem B. 2024; 128(29):6975-6989.

PMID: 38991095 PMC: 11821374. DOI: 10.1021/acs.jpcb.4c01277.

Innovative Approaches for Drug Discovery: Quantifying Drug Distribution and Response with Raman Imaging.

Dunnington E, Wong B, Fu D Anal Chem. 2024; 96(20):7926-7944.

PMID: 38625100 PMC: 11108735. DOI: 10.1021/acs.analchem.4c01413.

Accuracy of Raman spectroscopy in the diagnosis of Alzheimer's disease.

Xu Y, Pan X, Li H, Cao Q, Xu F, Zhang J Front Psychiatry. 2023; 14:1112615.

PMID: 37009107 PMC: 10060832. DOI: 10.3389/fpsyt.2023.1112615.

Computational conjugate adaptive optics microscopy for longitudinal through-skull imaging of cortical myelin.

Kwon Y, Hong J, Kang S, Lee H, Jo Y, Kim K Nat Commun. 2023; 14(1):105.

PMID: 36609405 PMC: 9823103. DOI: 10.1038/s41467-022-35738-9.

References

Raichle M, Gusnard D . Appraising the brain's energy budget. Proc Natl Acad Sci U S A. 2002; 99(16):10237-9. PMC: 124895. DOI: 10.1073/pnas.172399499. View

Navabpour S, Kwapis J, Jarome T . A neuroscientist's guide to transgenic mice and other genetic tools. Neurosci Biobehav Rev. 2019; 108:732-748. PMC: 8049509. DOI: 10.1016/j.neubiorev.2019.12.013. View

Rasmussen R, Nedergaard M, Petersen N . Sulforhodamine 101, a widely used astrocyte marker, can induce cortical seizure-like activity at concentrations commonly used. Sci Rep. 2016; 6:30433. PMC: 4960645. DOI: 10.1038/srep30433. View

Witte S, Negrean A, Lodder J, de Kock C, Testa Silva G, Mansvelder H . Label-free live brain imaging and targeted patching with third-harmonic generation microscopy. Proc Natl Acad Sci U S A. 2011; 108(15):5970-5. PMC: 3076839. DOI: 10.1073/pnas.1018743108. View

Evans C, Potma E, Puorishaag M, Cote D, Lin C, Xie X . Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. Proc Natl Acad Sci U S A. 2005; 102(46):16807-12. PMC: 1283840. DOI: 10.1073/pnas.0508282102. View

Fu D, Lu F, Zhang X, Freudiger C, Pernik D, Holtom G . Quantitative chemical imaging with multiplex stimulated Raman scattering microscopy. J Am Chem Soc. 2012; 134(8):3623-6. PMC: 3396204. DOI: 10.1021/ja210081h. View

Ji M, Orringer D, Freudiger C, Ramkissoon S, Liu X, Lau D . Rapid, label-free detection of brain tumors with stimulated Raman scattering microscopy. Sci Transl Med. 2013; 5(201):201ra119. PMC: 3806096. DOI: 10.1126/scitranslmed.3005954. View

Lu F, Basu S, Igras V, Hoang M, Ji M, Fu D . Label-free DNA imaging in vivo with stimulated Raman scattering microscopy. Proc Natl Acad Sci U S A. 2015; 112(37):11624-9. PMC: 4577158. DOI: 10.1073/pnas.1515121112. View

Hill A, Fu D . Cellular Imaging Using Stimulated Raman Scattering Microscopy. Anal Chem. 2019; 91(15):9333-9342. DOI: 10.1021/acs.analchem.9b02095. View

10.

Hollon T, Pandian B, Adapa A, Urias E, Save A, Khalsa S . Near real-time intraoperative brain tumor diagnosis using stimulated Raman histology and deep neural networks. Nat Med. 2020; 26(1):52-58. PMC: 6960329. DOI: 10.1038/s41591-019-0715-9. View

11.

Francis A, Berry K, Chen Y, Figueroa B, Fu D . Label-free pathology by spectrally sliced femtosecond stimulated Raman scattering (SRS) microscopy. PLoS One. 2017; 12(5):e0178750. PMC: 5451135. DOI: 10.1371/journal.pone.0178750. View

12.

Wei M, Shi L, Shen Y, Zhao Z, Guzman A, Kaufman L . Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy. Proc Natl Acad Sci U S A. 2019; 116(14):6608-6617. PMC: 6452712. DOI: 10.1073/pnas.1813044116. View

13.

Chen Y, Liu S, Liu H, Tong S, Tang H, Zhang C . Coherent Raman Scattering Unravelling Mechanisms Underlying Skull Optical Clearing for Through-Skull Brain Imaging. Anal Chem. 2019; 91(15):9371-9375. DOI: 10.1021/acs.analchem.9b02624. View

14.

Manifold B, Thomas E, Francis A, Hill A, Fu D . Denoising of stimulated Raman scattering microscopy images via deep learning. Biomed Opt Express. 2019; 10(8):3860-3874. PMC: 6701518. DOI: 10.1364/BOE.10.003860. View

15.

Hill A, Manifold B, Fu D . Tissue imaging depth limit of stimulated Raman scattering microscopy. Biomed Opt Express. 2020; 11(2):762-774. PMC: 7041472. DOI: 10.1364/BOE.382396. View

16.

Dong P, Lin H, Huang K, Cheng J . Label-free quantitation of glycated hemoglobin in single red blood cells by transient absorption microscopy and phasor analysis. Sci Adv. 2019; 5(5):eaav0561. PMC: 6510558. DOI: 10.1126/sciadv.aav0561. View

17.

Francis A, Shears M, Murphy S, Fu D . Direct Quantification of Single Red Blood Cell Hemoglobin Concentration with Multiphoton Microscopy. Anal Chem. 2020; 92(18):12235-12241. DOI: 10.1021/acs.analchem.0c01609. View

18.

Chen T, Huang Y . Label-Free Transient Absorption Microscopy for Red Blood Cell Flow Velocity Measurement in Vivo. Anal Chem. 2017; 89(19):10120-10123. DOI: 10.1021/acs.analchem.7b01952. View

19.

Snaidero N, Simons M . Myelination at a glance. J Cell Sci. 2014; 127(Pt 14):2999-3004. DOI: 10.1242/jcs.151043. View

20.

Tian F, Yang W, Mordes D, Wang J, Salameh J, Mok J . Monitoring peripheral nerve degeneration in ALS by label-free stimulated Raman scattering imaging. Nat Commun. 2016; 7:13283. PMC: 5095598. DOI: 10.1038/ncomms13283. View