The cranial and spinal windows for optical neuroimaging in vivo
| Subject | Animal | Mouse | Mouse | Mouse | Mouse | Mouse | Mouse | Mouse | Mouse |
|---|---|---|---|---|---|---|---|---|---|
| Species | Male B6CBAF1 or female Thy1.2YFP-H | YFP-H, CX3CR1-GFP, CX3CR1-GFP x YFP-H, Emx-1-cre, GFAP-GFP | Thy1-YFP-H and GFP-M (neuron) Balb/c and C57BL/6 wildtype (glioma) |
Female transgenic C57BL/6 expressing GCaMP6s/f and GFP-m | C57BL/6 male | C57BL/6, Ai95(RCL-GCaMP6f)-D, NG2-CreERT2, Thy1-GCaMP6f | Sox9 cKO: Sox9fl/fl; CAG-CreER; Aldh1L1-EGFP Control: Sox9fl/fl; Aldh1L1-EGFP |
G7NG817, Thy1-EYFP-H | |
| Anesthetization | Awake | Anesthetized with isoflurane | Anesthetized with isoflurane | Awake | Anesthetized with isoflurane | Awake | Anesthetized with isoflurane | Awake or anesthetized with isoflurane | |
| Region of interest | Primary sensory cortical areas (S1HL |
Spinal cord: vertebrae (T10-T12 |
CA1 |
Sensory and motor cortex | Primary sensory cortical areas (S1FL |
Olfactory bulb | Olfactory bulb | Primary visual cortex (V1 |
|
| Window | Window type | Open-close window (plug) | Open-close window (plug) | Modified sealed cranial window | Thinned-skull or intact skull window | Sealed cranial window | Sealed cranial window | Thinned-skull window | PEO-CYTOP |
| Window size | ~4 mm in diameter | 5 mm in diameter | 0.84 mm in diameter | ~ 9×9 mm | 6 mm in diameter | 2~3 mm in diameter | > 8 mm in diameter | ||
| Window material | Glass coverslip | Metal bars, glass coverslip | Glass capillary, glass coverslip, stainless steel screws | Glass coverslip, steel head bar | Anodized titanium | A titanium head-bar, glass coverslip | Glass coverslip | PEO-CYTOP nanosheet | |
| Bonding material | N/A | Dental acrylic and cyanoacrylate | Dental acrylic, UV curing epoxy | Mixture of metabond and dental cement | Dental cement | Photopolymerizable dental cement | N/A | UV curable dental acrylic | |
| Optical imaging | Imaging technique | 2PE-CaSDI | 2PEF | 1PEF and 2PEF | OISI | OISI, LSCI | 2PE-CaSDI | 2PE-CaSDI | 2PEF, 2PE-CaSDI |
| Measuring frequency (temporal resolution) | (256 msec/frame) | N/A | 100 Hz (up to 1.2 kHz) | 30 Hz | 10 Hz/5 Hz (200 msec/ 400 msec) | (50~150 msec/frame) | 10~35 Hz | 1 Hz for EYFP, 30 Hz for GCaMP7, 3.8 Hz (CaSDI) | |
| Measuring time | 4~5 min | N/A | Multiple sessions (30~60 min) | 30 sec or 11.5 and 5 min | 20 sec | N/A | 10 sec/trial | N/A | |
| Objective lens | 40X/0.8 NA |
20X/1.0 NA water-immersion, 40X/0.8 NA, water-immersion, 4X/0.28 NA | 10X/0.25 NA, 20X/0.40 NA (1PEF) 10X/0.25 NA (2PEF) |
Adjustable lens (f |
4X/0.10 NA | 60X/1.10NA, 40X/0.8NA | 20X/1.0 NA | 16X/0.80 NA water-immersion (cross-sectional imaging), 25X/1.10 NA water-immersion (deep in vivo imaging), 2X/0.20NA (CaSDI) | |
| Test & Apparatus | Animal test | Spherical treadmill running training | Open field and runway assays | N/A | Automated self-head-fixation and visual-evoked cortical imaging | Somatosensory-evoked functional imaging | Synaptic activity triggers odor-specific OPC |
Three compartment place preference assay | CaSDI in vast FOV, deep and wide-field imaging |
| Behavior test apparatus | Air-supported free-floating Styrofoam ball | Plexiglass enclosure to enter a dark goal box at the end of the runway | N/A | Fully automated and self-initiated head-fixation system for functional imaging, yellow light flash stimulator | Electrical sensory stimulator | Sensory stimulation, running wheel with head fixed | A testing chamber, video with a behavior tracking camera (for odor discrimination) | N/A | |
| Life supporting system | N/A | Rectal thermometer and feedback-controlled heating blanket at 37.5°C | Heating blanket | N/A | 37°C heating pad | 36.5~37°C heating pad, pneumogram transducer | Heating pad | Disposable heating pad | |
| Ref. | Published year | 2007 | 2012 | 2011 | 2016 | 2016 | 2018 | 2021 | 2020, 2021 |
| First author | Dombeck, D. A. | Farrar, M. J. | Barretto, R. P. J. | Murphy, T. H. | Cho, A. | Rungta, R. L. | Ung, K. | Takahashi, T. | |
The cranial and spinal window types were varied by the research objectives, animal condition and optical imaging methods.
aS1HL primary somatosensory hindlimb cortex; bT10-T12 tenth to twelfth thoracic vertebra; cCA1 the first region in the hippocampal circuit; dS1FL primary somatosensory forelimb cortex; eV1 primary visual cortex; fNA numerical aperture; gf focal length; hOPC oligodendrocyte precursor cell.