Raman spectroscopy in microfluidic chips reveals hyphal scale stress-associated metabolic responses in filamentous soil fungi
Pucetaite, Milda LU ; Mafla-Endara, Paola M. LU ; González, Yitsully G. ; Zou, Hanbang LU ; Schmidt, Robert W. ; Ariese, Freek and Hammer, Edith C. LU
(2026)
In Fungal Biology
130(3).
- Abstract
Understanding metabolic processes of soil fungi is essential for elucidating their ecological roles in biogeochemical cycles and responses to emergent environmental stressors. Here, we demonstrate the potential of using stable isotope probing Raman (SIP-Raman) microspectroscopy in microfluidics technology-based soil chips to trace glucose metabolism rates and stress responses in laboratory grown filamentous soil fungus Psilocybe cf. subviscida . The time evolution of Raman spectral band intensities resulting from deuterated glucose uptake in the fungal hyphae allowed us to assess glucose metabolism rates. Under excess copper (Cu) stress, we observed suppression of both glucose metabolic activity and growth. In addition, reduced spectral... (More)
Understanding metabolic processes of soil fungi is essential for elucidating their ecological roles in biogeochemical cycles and responses to emergent environmental stressors. Here, we demonstrate the potential of using stable isotope probing Raman (SIP-Raman) microspectroscopy in microfluidics technology-based soil chips to trace glucose metabolism rates and stress responses in laboratory grown filamentous soil fungus Psilocybe cf. subviscida . The time evolution of Raman spectral band intensities resulting from deuterated glucose uptake in the fungal hyphae allowed us to assess glucose metabolism rates. Under excess copper (Cu) stress, we observed suppression of both glucose metabolic activity and growth. In addition, reduced spectral signatures of intracellular cytochrome c further implied impaired mitochondrial function and potential onset of cell death. However, laser-induced radiation damage hampered repeated Raman measurements, including multispectral mapping, on individual hyphae, especially when exposed to the Cu stress. To overcome this, we employed stimulated Raman scattering (SRS) microscopy, which offers much higher sensitivity and mapping speeds, and therefore much lower radiation doses. This enabled localization of the uptaken glucose at the inner edges of the P. cf. subviscida hyphae and Cu-induced formation of putative vacuolar structures. While integration of this approach with soil chips requires future modifications to the chip design for increased optical transparency and ensured sterility, overall, our results demonstrate the potential of Raman-based microspectroscopy for spatially resolved, in situ analysis of fungal primary metabolism and stress physiology.
(Less)
- author
- Pucetaite, Milda
LU
; Mafla-Endara, Paola M.
LU
; González, Yitsully G.
; Zou, Hanbang
LU
; Schmidt, Robert W.
; Ariese, Freek
and Hammer, Edith C.
LU
- organization
- publishing date
- 2026-05
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- metabolic activity, Microfluidic chips, Raman scattering microspectroscopy, Soil fungi, Stable-isotope probing, Stimulated Raman scattering (SRS) microscopy
- in
- Fungal Biology
- volume
- 130
- issue
- 3
- article number
- 101749
- publisher
- Elsevier
- external identifiers
-
- scopus:105032891839
- ISSN
- 1878-6146
- DOI
- 10.1016/j.funbio.2026.101749
- project
- Using stable-isotope vibrational microspectroscopy to reveal hyphal scale interactions and competition between ectomycorrhizal and saprotrophic fungi
- language
- English
- LU publication?
- yes
- additional info
- Publisher Copyright: © 2026 The Authors.
- id
- 3cb61cb8-c89e-49c5-8c09-5237ed659b6e
- date added to LUP
- 2026-04-02 11:36:02
- date last changed
- 2026-04-10 14:38:12
@article{3cb61cb8-c89e-49c5-8c09-5237ed659b6e,
abstract = {{<p>Understanding metabolic processes of soil fungi is essential for elucidating their ecological roles in biogeochemical cycles and responses to emergent environmental stressors. Here, we demonstrate the potential of using stable isotope probing Raman (SIP-Raman) microspectroscopy in microfluidics technology-based soil chips to trace glucose metabolism rates and stress responses in laboratory grown filamentous soil fungus Psilocybe cf. subviscida . The time evolution of Raman spectral band intensities resulting from deuterated glucose uptake in the fungal hyphae allowed us to assess glucose metabolism rates. Under excess copper (Cu) stress, we observed suppression of both glucose metabolic activity and growth. In addition, reduced spectral signatures of intracellular cytochrome c further implied impaired mitochondrial function and potential onset of cell death. However, laser-induced radiation damage hampered repeated Raman measurements, including multispectral mapping, on individual hyphae, especially when exposed to the Cu stress. To overcome this, we employed stimulated Raman scattering (SRS) microscopy, which offers much higher sensitivity and mapping speeds, and therefore much lower radiation doses. This enabled localization of the uptaken glucose at the inner edges of the P. cf. subviscida hyphae and Cu-induced formation of putative vacuolar structures. While integration of this approach with soil chips requires future modifications to the chip design for increased optical transparency and ensured sterility, overall, our results demonstrate the potential of Raman-based microspectroscopy for spatially resolved, in situ analysis of fungal primary metabolism and stress physiology.</p>}},
author = {{Pucetaite, Milda and Mafla-Endara, Paola M. and González, Yitsully G. and Zou, Hanbang and Schmidt, Robert W. and Ariese, Freek and Hammer, Edith C.}},
issn = {{1878-6146}},
keywords = {{metabolic activity; Microfluidic chips; Raman scattering microspectroscopy; Soil fungi; Stable-isotope probing; Stimulated Raman scattering (SRS) microscopy}},
language = {{eng}},
number = {{3}},
publisher = {{Elsevier}},
series = {{Fungal Biology}},
title = {{Raman spectroscopy in microfluidic chips reveals hyphal scale stress-associated metabolic responses in filamentous soil fungi}},
url = {{http://dx.doi.org/10.1016/j.funbio.2026.101749}},
doi = {{10.1016/j.funbio.2026.101749}},
volume = {{130}},
year = {{2026}},
}