Label-free visualization of cholesteatoma in the mastoid and tympanic membrane using CARS microscopy

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• 作者：Jing Zou^a ; ^b ; ^{Jing.Zou@uta.fi" class="auth_mail" title="E-mail the corresponding author} ; ^{zoujinghb@hotmail.com" class="auth_mail" title="E-mail the corresponding author} ; Antti Isomä ; ki^d ; Timo Hirvonen^c ; Antti Aarnisalo^c ; Jussi Jero^c ; Ilmari Pyykkö ; ^b
• 关键词：Cholesteatoma ; Imaging ; Label-free ; Raman spectroscopy ; High resolution
• 刊名：Journal of Otology
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：11
• 期：3
• 页码：127-133
• 全文大小：2362 K

文摘

The present study aimed to evaluate the possibility of using coherent anti-Stokes Raman spectroscopy (CARS) microscopy to determine the specific molecular morphology of cholesteatoma by detecting the natural vibrational contrast of the chemical bonds without any staining.

Materials and methods

Specimens from the mastoid and tympanic membrane with and without cholesteatoma were analyzed using CARS microscopy, two-photon excited fluorescence (TPEF) microscopy, and the second harmonic generation (SHG) microscopy.

Results

In cholesteatoma tissues from the mastoid, a strong resonant signal at 2845 cm⁻¹ was observed by CARS, which indicated the detection of the CH₂ hydro-carbon lipid bonds that do not generate visible signals at 2940 cm⁻¹ suggestive of CH₃ bonds in amino acids. A strong resonant signal at 2940 cm⁻¹ appeared in an area of the same specimen, which also generated abundant signals by TPEF and SHG microscopy at 817 nm, which was suggestive of collagen. In the tympanic membrane specimen with cholesteatoma, a strong resonant signal with corrugated morphology was detected, which indicated the presence of lipids. A strong signal was detected in the tympanic membrane with chronic otitis media using TPEF/SHG at 817 nm, which indicated collagen enrichment. The CARS and TPEF/SHG images were in accordance with the histology results.

Conclusion

These results suggest the need to develop a novel CARS microendoscope that can be used in combination with TPEF/SHG to distinguish cholesteatoma from inflammatory tissues.