Label-free superior contrast with c-band ultra-violet extinction microscopy

Florian Ströhl; Deanna L. Wolfson; Ida S. Opstad; Daniel H. Hansen; Hong Mao; Balpreet S. Ahluwalia

doi:10.1038/s41377-023-01105-6

Go to Nature Website

Your Location：

Home >

Browse articles >

Label-free superior contrast with c-band ultra-violet extinction microscopy

Original Articles

- Label-free superior contrast with c-band ultra-violet extinction microscopy
- Light: Science & Applications Vol. 12, Issue 3, Pages: 435-445(2023)
- Affiliations：
  
  1.Department of Physics and Technology, UiT The Arctic University of Norway, Tromsø, Norway
  2.Department of Clinical Science, Intervention and Technology, Karolinska Institute, Stockholm, Sweden
- Author bio：
  
  Florian Ströhl (florian.strohl@uit.no)
- Funds：
- DOI：10.1038/s41377-023-01105-6
  CLC：
- Published：31 March 2023，
  
  Published Online：3 March 2023，
  
  Received：27 October 2022，
  
  Revised：14 February 2023，
  
  Accepted：14 February 2023
Scan for full text
Ströhl, F. et al. Label-free superior contrast with c-band ultra-violet extinction microscopy. Light: Science & Applications, 12, 435-445 (2023).
DOI：

Ströhl, F. et al. Label-free superior contrast with c-band ultra-violet extinction microscopy. Light: Science & Applications, 12, 435-445 (2023). DOI： 10.1038/s41377-023-01105-6.

摘要

Abstract

In 1934

Frits Zernike demonstrated that it is possible to exploit the sample's refractive index to obtain superior contrast images of biological cells. The refractive index contrast of a cell surrounded by media yields a change in the phase and intensity of the transmitted light wave. This change can be due to either scattering or absorption caused by the sample. Most cells are transparent at visible wavelengths

which means the imaginary component of their complex refractive index

also known as extinction coefficient

is close to zero. Here

we explore the use of c-band ultra-violet (UVC) light for high-contrast high-resolution label-free microscopy

as k is naturally substantially higher in the UVC than at visible wavelengths. Using differential phase contrast illumination and associated processing

we achieve a 7- to 300-fold improvement in contrast compared to visible-wavelength and UVA differential interference contrast microscopy or holotomography

and quantify the extinction coefficient distribution within liver sinusoidal endothelial cells. With a resolution down to 215 nm

we are

for the first time in a far-field label-free method

able to image individual fenestrations within their sieve plates which normally requires electron or fluorescence superresolution microscopy. UVC illumination also matches the excitation peak of intrinsically fluorescent proteins and amino acids and thus allows us to utilize autofluorescence as an independent imaging modality on the same setup.

关键词

Keywords

references

Popescu, G. Quantitative Phase Imaging of Cells and Tissues (McGraw-Hill, 2011).

Kubitscheck, U. Fluorescence Microscopy: From Principles to Biological Applications, 2nd edn. (Wiley-VCH Verlag GmbH & Co. KGaA, 2017).

Y. Cotte , 等 . Marker-free phase nanoscopy . Nat. Photonics , 2013 . 7 113 -117 . DOI:10.1038/nphoton.2012.329http://doi.org/10.1038/nphoton.2012.329 .

Popescu, G. Nanobiophotonics (McGraw-Hill, 2010).

S. B. Mehta , C. J. R. Sheppard . Quantitative phase-gradient imaging at high resolution with asymmetric illumination-based differential phase contrast . Opt. Lett. , 2009 . 34 1924 DOI:10.1364/ol.34.001924http://doi.org/10.1364/ol.34.001924 .

M. Chen , Z. F. Phillips , L. Waller . Quantitative differential phase contrast (DPC) microscopy with computational aberration correction . Opt. Express , 2018 . 26 32888 DOI:10.1364/oe.26.032888http://doi.org/10.1364/oe.26.032888 .

G. Zheng , R. Horstmeyer , C. Yang , G. Zheng , C. Yang . Wide-field, high-resolution Fourier ptychographic microscopy . Nat. Photonics , 2013 . 7 739 -745 . DOI:10.1038/nphoton.2013.187http://doi.org/10.1038/nphoton.2013.187 .

A. Ojaghi , 等 . Label-free hematology analysis using deep-ultraviolet microscopy . Proc. Natl Acad. Sci. USA , 2020 . 117 14779 -14789 . DOI:10.1073/pnas.2001404117http://doi.org/10.1073/pnas.2001404117 .

F. Fereidouni , 等 . Microscopy with ultraviolet surface excitation for rapid slide-free histology . Nat. Biomed. Eng. , 2017 . 1 957 -966 . DOI:10.1038/s41551-017-0165-yhttp://doi.org/10.1038/s41551-017-0165-y .

S. Klinke , 等 . Crystallization and preliminary X-ray characterization of the full-length bacteriophytochrome from the plant pathogen Xanthomonas campestris pv. campestris . Acta Crystallogr. F: Struct. Biol. Commun. , 2014 . 70 1636 -1639 . DOI:10.1107/S2053230X14023243http://doi.org/10.1107/S2053230X14023243 .

S. Klinke , M. J. Suárez , A. González-Pérez , J. M. Ruso , F. Sarmiento . A spectroscopic study of the interaction catalase-cationic surfactant (n-decyltrimethylammonium bromide) in aqueous solutions at different pH and temperatures . Phys. Chem. Chem. Phys. , 2004 . 6 816 -821 . DOI:10.1039/B308466Chttp://doi.org/10.1039/B308466C .

T. Inagaki , R. N. Hamm , E. T. Arakawa , L. R. Painter . Optical and dielectric properties of DNA in the extreme ultraviolet . J. Chem. Phys. , 1974 . 61 4246 -4250 . DOI:10.1063/1.1681724http://doi.org/10.1063/1.1681724 .

S. Kaščáková , 等 . Ex vivo quantification of mTHPC concentration in tissue: influence of chemical extraction on the optical properties . J. Photochem. Photobiol. B: Biol. , 2008 . 91 99 -107 . DOI:10.1016/j.jphotobiol.2008.02.003http://doi.org/10.1016/j.jphotobiol.2008.02.003 .

A. Matlock , A. Sentenac , P. C. Chaumet , J. Yi , L. Tian . Inverse scattering for reflection intensity phase microscopy . Biomed. Opt. Express , 2020 . 11 911 DOI:10.1364/boe.380845http://doi.org/10.1364/boe.380845 .

A. Faridian , 等 . Nanoscale imaging using deep ultraviolet digital holographic microscopy . Opt. Express , 2010 . 18 14159 DOI:10.1364/oe.18.014159http://doi.org/10.1364/oe.18.014159 .

A. K. Singh , A. Faridian , P. Gao , G. Pedrini , W. Osten . Quantitative phase imaging using a deep UV LED source . Opt. Lett. , 2014 . 39 3468 DOI:10.1364/ol.39.003468http://doi.org/10.1364/ol.39.003468 .

Y. Zhang , H. Gross . Systematic design of microscope objectives. I: System review and analysis . Adv. Opt. Technol. , 2019 . 8 DOI:10.1515/aot-2019-0013http://doi.org/10.1515/aot-2019-0013 .

W. Vollrath . Ultra-high-resolution DUV microscope optics for semiconductor applications . Proc. SPIE , 2005 . 5865 58650E DOI:10.1117/12.624560http://doi.org/10.1117/12.624560 .

A. Descloux , K. S. Grußmayer , A. Radenovic . Parameter-free image resolution estimation based on decorrelation analysis . Nat. Methods , 2019 . 16 918 -924 . DOI:10.1038/s41592-019-0515-7http://doi.org/10.1038/s41592-019-0515-7 .

R. Fraser , 等 . The liver sieve and atherosclerosis . Pathology , 2012 . 44 181 -186 . DOI:10.1097/PAT.0b013e328351bcc8http://doi.org/10.1097/PAT.0b013e328351bcc8 .

F. Ströhl , C. F. Kaminski . Frontiers in structured illumination microscopy . Optica , 2016 . 3 667 DOI:10.1364/OPTICA.3.000667http://doi.org/10.1364/OPTICA.3.000667 .

V. C. Cogger , 等 . Three-dimensional structured illumination microscopy of liver sinusoidal endothelial cell fenestrations . J. Struct. Biol. , 2010 . 171 382 -388 . DOI:10.1016/j.jsb.2010.06.001http://doi.org/10.1016/j.jsb.2010.06.001 .

E. Peli . Contrast in complex images . J. Opt. Soc. Am. A , 1990 . 7 2032 DOI:10.1364/JOSAA.7.002032http://doi.org/10.1364/JOSAA.7.002032 .

J. Ortega-Arroyo , P. Kukura . Interferometric scattering microscopy (iSCAT): new frontiers in ultrafast and ultrasensitive optical microscopy . Phys. Chem. Chem. Phys. , 2012 . 14 15625 DOI:10.1039/c2cp41013chttp://doi.org/10.1039/c2cp41013c .

I. Arganda-Carreras , 等 . Trainable Weka Segmentation: a machine learning tool for microscopy pixel classification . Bioinformatics , 2017 . 33 2424 -2426 . DOI:10.1093/bioinformatics/btx180http://doi.org/10.1093/bioinformatics/btx180 .

V. Mönkemöller , C. Øie , W. Hübner , T. Huser , P. McCourt . Multimodal super-resolution optical microscopy visualizes the close connection between membrane and the cytoskeleton in liver sinusoidal endothelial cell fenestrations . Sci. Rep. , 2015 . 5 1 -10 . DOI:10.1038/srep16279http://doi.org/10.1038/srep16279 .

C. I. Øie , 等 . New ways of looking at very small holes—using optical nanoscopy to visualize liver sinusoidal endothelial cell fenestrations . Nanophotonics , 2018 . 7 575 -596 . DOI:10.1515/nanoph-2017-0055http://doi.org/10.1515/nanoph-2017-0055 .

T. A. Smith , L. M. Hirvonen , C. N. Lincoln , X. Hao . Deep-UV confocal fluorescence imaging and super-resolution optical microscopy of biological samples . J. Innov. Opt. Health Sci. , 2012 . 5 1 -9 . DOI:10.1142/S1793545812500253http://doi.org/10.1142/S1793545812500253 .

F. Jamme , 等 . Deep UV autofluorescence microscopy for cell biology and tissue histology . Biol. Cell , 2013 . 105 277 -288 . DOI:10.1111/boc.201200075http://doi.org/10.1111/boc.201200075 .

P. A. Heimann , R. Urstadt . Deep ultraviolet microscope . Appl. Opt. , 1990 . 29 495 DOI:10.1364/ao.29.000495http://doi.org/10.1364/ao.29.000495 .

A. B. T. Ghisaidoobe , S. J. Chung . Intrinsic tryptophan fluorescence in the detection and analysis of proteins: A focus on förster resonance energy transfer techniques . Int. J. Mol. Sci. , 2014 . 15 22518 -22538 . DOI:10.3390/ijms151222518http://doi.org/10.3390/ijms151222518 .

C. S. Downes , A. R. Collins , R. T. Johnson . DNA damage in synchronized HeLa cells irradiated with ultraviolet . Biophys. J. , 1979 . 25 129 -150 . DOI:10.1016/S0006-3495(79)85282-0http://doi.org/10.1016/S0006-3495(79)85282-0 .

J. H. Stockley , 等 . Surpassing light-induced cell damage in vitro with novel cell culture media . Sci. Rep. , 2017 . 7 1 -11 . DOI:10.1038/s41598-017-00829-xhttp://doi.org/10.1038/s41598-017-00829-x .

B. Moulden , F. Kingdom , L. F. Gatley . The standard deviation of luminance as a metric for contrast in random-dot images . Perception , 1990 . 19 79 -101 . DOI:10.1068/p190079http://doi.org/10.1068/p190079 .

F. Ströhl , S. Jadhav , B. Ahluwalia , K. Agarwal , D. Prasad . Object detection neural network improves Fourier ptychography reconstruction . Opt. Express , 2020 . 28 37199 -37208 . DOI:10.1364/oe.409679http://doi.org/10.1364/oe.409679 .

J. Schindelin , 等 . Fiji: an open-source platform for biological-image analysis . Nat. Methods , 2012 . 9 676 -682 . DOI:10.1038/nmeth.2019http://doi.org/10.1038/nmeth.2019 .

B. Smedsrød , H. Pertoft , G. Eggertsen , C. Sundström . Functional and morphological characterization of cultures of Kupffer cells and liver endothelial cells prepared by means of density separation in Percoll, and selective substrate adherence . Cell Tissue Res. , 1985 . 241 639 -649 . DOI:10.1007/BF00214586http://doi.org/10.1007/BF00214586 .

V. Mönkemöller , 等 . Primary rat LSECs preserve their characteristic phenotype after cryopreservation . Sci. Rep. , 2018 . 8 1 -10 . DOI:10.1038/s41598-018-32103-zhttp://doi.org/10.1038/s41598-018-32103-z .

Views

Downloads

CSCD

Alert me when the article has been cited

Submit

Tools

Publicity Resources

No data

Related Author

No data

Related Institution

No data

⁰