Label-free third harmonic generation imaging and quantification of lipid droplets in live filamentous fungi
2022
Authors:
Pajić, TanjaTodorović, Nataša
Živić, Miroslav
Nikolić, Stanko N
Rabasović, Mihailo D
Clayton, Andrew H A
Krmpot, Aleksandar J
Document Type:
Article (Published version)
Metadata
Show full item recordAbstract:
We report the utilization of Third-Harmonic Generation microscopy for label-free live cell imaging of lipid droplets in the hypha of filamentous fungus Phycomyces blakesleeanus . THG microscopy images showed bright spherical features dispersed throughout the hypha cytoplasm in control conditions and a transient increase in the number of bright features after complete nitrogen starvation. Colocalization analysis of THG and lipid-counterstained images disclosed that the cytoplasmic particles were lipid droplets. Particle Size Analysis and Image Correlation Spectroscopy were used to quantify the number density and size of lipid droplets. The two analysis methods both revealed an increase from 16 × 10 −3 to 23 × 10 −3 lipid droplets/µm 2 after nitrogen starvation and a decrease in the average size of the droplets (range: 0.5–0.8 µm diameter). In conclusion, THG imaging, followed by PSA and ICS, can be reliably used for filamentous fungi for the in vivo quantification of lipid droplets without the need for labeling and/or fixation. In addition, it has been demonstrated that ICS is suitable for THG microscopy.
Source:
Scientific Reports, 2022, 12, 1, 18760-Funding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200007 (University of Belgrade, Institute for Biological Research 'Siniša Stanković') (RS-MESTD-inst-2020-200007)
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200178 (University of Belgrade, Faculty of Biology) (RS-MESTD-inst-2020-200178)
- HEMMAGINERO - Hemoglobin-Based Spectroscopy and Nonlinear Imaging of Erythrocytes and Their Membranes As Emerging Diagnostic Tool (RS-ScienceFundRS-Promis-6066079)
DOI: 10.1038/s41598-022-23502-4
ISSN: 2045-2322
PubMed: 36335164
WoS: 000879722100061
Scopus: 2-s2.0-85141401657
URI
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC9637149https://www.nature.com/articles/s41598-022-23502-4
http://radar.ibiss.bg.ac.rs/handle/123456789/5235