Compressive hyperspectral time-resolved wide-field fluorescence lifetime imaging

AbstractA deeper understanding of spatial resolution in microscopy fostered a technological revolution that is now permitting us to investigate the structure of the cell with nanometer resolution. Although fluorescence microscopy techniques enable scientists to investigate both the structure and biochemistry of the cell, the biochemical resolving power of a microscope is a physical quantity that is not well-defined or studied. To overcome this limitation, we carried out a theoretical investigation of the biochemical resolving power in fluorescence lifetime imaging microscopy, one of the most effective tools to investigate biochemistry in single living cells. With the theoretical analysis of information theory and Monte Carlo simulations, we describe how the ‘biochemical resolving power’ in time-resolved sensing depends on instrument specifications. We unravel common misunderstandings on the role of the instrument response function and provide theoretical insights that have significant practical implications in the design and use of time-resolved instrumentation.

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Mapping the Spatiotemporal Heterogeneity of Biomolecules Concentration, Mobility and Local Environment in Live Cells using Quantitative Time-Resolved Confocal Fluorescence Microscopy Imaging Without Scanning and Fluorescence Lifetime Imaging Microscopy

Biophysical Journal ◽

10.1016/j.bpj.2019.11.1735 ◽

2020 ◽

Vol 118 (3) ◽

pp. 307a

Author(s):

Sho Oasa ◽

Aleksandar Krmpot ◽

Stanko Nikolic ◽

Lars Terenius ◽

Rudolf Rigler ◽

...

Keyword(s):

Fluorescence Microscopy ◽

Fluorescence Lifetime ◽

Local Environment ◽

Live Cells ◽

Fluorescence Lifetime Imaging Microscopy ◽

Fluorescence Lifetime Imaging ◽

Microscopy Imaging ◽

Time Resolved ◽

Lifetime Imaging ◽

Confocal Fluorescence

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Macroscopic fluorescence-lifetime imaging of NADH and protoporphyrin IX improves the detection and grading of 5-aminolevulinic acid-stained brain tumors

Scientific Reports ◽

10.1038/s41598-020-77268-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Mikael T. Erkkilä ◽

David Reichert ◽

Johanna Gesperger ◽

Barbara Kiesel ◽

Thomas Roetzer ◽

...

Keyword(s):

Brain Tumor ◽

Brain Tumors ◽

Fluorescence Lifetime ◽

Tumor Tissue ◽

Aminolevulinic Acid ◽

Protoporphyrin Ix ◽

Low Grade ◽

Wide Field ◽

Fluorescence Lifetime Imaging ◽

Lifetime Imaging

AbstractMaximal safe tumor resection remains the key prognostic factor for improved prognosis in brain tumor patients. Despite 5-aminolevulinic acid-based fluorescence guidance the neurosurgeon is, however, not able to visualize most low-grade gliomas (LGG) and infiltration zone of high-grade gliomas (HGG). To overcome the need for a more sensitive visualization, we investigated the potential of macroscopic, wide-field fluorescence lifetime imaging of nicotinamide adenine dinucleotide (NADH) and protoporphyrin IX (PPIX) in selected human brain tumors. For future intraoperative use, the imaging system offered a square field of view of 11 mm at 250 mm free working distance. We performed imaging of tumor tissue ex vivo, including LGG and HGG as well as brain metastases obtained from 21 patients undergoing fluorescence-guided surgery. Half of all samples showed visible fluorescence during surgery, which was associated with significant increase in PPIX fluorescence lifetime. While the PPIX lifetime was significantly different between specific tumor tissue types, the NADH lifetimes did not differ significantly among them. However, mainly necrotic areas exhibited significantly lower NADH lifetimes compared to compact tumor in HGG. Our pilot study indicates that combined fluorescence lifetime imaging of NADH/PPIX represents a sensitive tool to visualize brain tumor tissue not detectable with conventional 5-ALA fluorescence.

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