Selective plane illumination microscopy (SPIM) with time-domain fluorescence lifetime imaging microscopy (FLIM) for volumetric measurement of cleared mouse brain samples.
Rev Sci Instrum
; 89(5): 053705, 2018 May.
Article
in En
| MEDLINE
| ID: mdl-29864842
ABSTRACT
We have developed an imaging technique which combines selective plane illumination microscopy with time-domain fluorescence lifetime imaging microscopy (SPIM-FLIM) for three-dimensional volumetric imaging of cleared mouse brains with micro- to mesoscopic resolution. The main features of the microscope include a wavelength-adjustable pulsed laser source (Tisapphire) (near-infrared) laser, a BiBO frequency-doubling photonic crystal, a liquid chamber, an electrically focus-tunable lens, a cuvette based sample holder, and an air (dry) objective lens. The performance of the system was evaluated with a lifetime reference dye and micro-bead phantom measurements. Intensity and lifetime maps of three-dimensional human embryonic kidney (HEK) cell culture samples and cleared mouse brain samples expressing green fluorescent protein (GFP) (donor only) and green and red fluorescent protein [positive Förster (fluorescence) resonance energy transfer] were acquired. The results show that the SPIM-FLIM system can be used for sample sizes ranging from single cells to whole mouse organs and can serve as a powerful tool for medical and biological research.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Brain
/
Imaging, Three-Dimensional
/
Microscopy
Limits:
Animals
/
Humans
/
Male
Language:
En
Journal:
Rev Sci Instrum
Year:
2018
Document type:
Article
Affiliation country:
United States