Table 1 Summary of optical sectioning techniques
From: Optical sectioning methods in three-dimensional bioimaging
Optical path setting | principle | Technique | Type | R*/ μm | SBR/dB | Through-Put*/ Mpx/s | Speed */ mm2/s | Advantages | Disadvantages | Applications |
|---|---|---|---|---|---|---|---|---|---|---|
Coaxial imaging | Focal plane conjugation | Confocal | Conventional191 | 0.22 | 12 | 7.9 | 0.16 | Flexible OS | Low scanning speed | Local fine imaging of biological tissue |
Deep learning32 | 0.12 | / | 12 | 0.17 | High throughput | Long training time | Organelle interactions | |||
ISM192 | 0.12 | / | 1.2 | 0.08 | High resolution | Low scanning speed | Cellular structures imaging | |||
Spinning disk193 | / | 0.51 | / | 419 | 2.8 | High throughput | Crosstalk | Intravital imaging | ||
Line confocal26 | / | 0.42 | 6.3 | 4915 | 27 | High scanning speed | Low OS | Whole slide imaging | ||
Intra-focal excitation | Two-photon | Conventional58 | 1.2 | 1 | 7.9 | 0.16 | Deep penetration | High light bleaching | ||
Multi-focus62 | 1.4 | / | 200 | 0.19 | High throughput | Severe tissue scattering | ||||
Line scanning69 | 0.77 | 5.5 | 98 | 8.5 | High throughput | Severe tissue scattering | Intravital imaging | |||
Wide field74 | 0.61 | / | 419 | 9.9 (10X) | Dynamic observation | |||||
TRIF80 | / | 0.21 | / | 419 | 0.11 (100X) | High axial resolution | Depth Limitation | Organelle interactions | ||
Modulated illumination | SIM | OS-SIM85 | 0.39 | 12 | 69 | 2.1 | Low phototoxicity | Illumination pattern sensitive | Large sample imaging | |
SR-SIM with PSF engineering94 | 0.09 | / | 140 | 0.09 (100X) | High resolution | Manual adjustment | Cellular structures imaging | |||
TRIF-SIM80 | 0.10 | / | 70 | 0.08 (100X) | High OS | Depth limitation | ||||
3D SIM96 | 0.10 | / | 28 | 0.05 (100X) | 3D super-resolution with high OS | Strict for optical alignment | ||||
Scanning SIM194 | 0.10 | / | / | / | High quality in modulation | Low scanning speed | ||||
Deep-learning SIM105 | 0.52 | 12 | 419 | 2.8 | Low noise levels | Large training data | Tissue imaging | |||
HiLo | Conventional86 | 0.41 | 19 | 209 | 2.6 | High temporal resolution | Long post-processing time | Intravital imaging | ||
Line-scanning HiLo116 | 7.5 | 25 | 23 | / | High scanning speed | Long post-processing time | ||||
Off-axis imaging | Mixed detection | Light sheet | SPIM195 | 0.68 | 3.7 | 25 | 1.4 | Low phototoxicity | Heterogeneous OS | Clear tissue imaging |
Bessel light sheet74 | 0.68 | / | 11 | 0.81 | Large thin illumination | Sample size limitation | ||||
iSPIM136 | 0.52 | / | 32 | 6.4 | Isotropic resolution | Objective limitation | Clear thick tissue imaging | |||
siSPIM153 | 0.45 | / | 248 | 17 | No size limitation | Low energy efficiency | Large, clear, thick tissue | |||
DSIM16 | / | 0.45 | / | 53 | 5.0 | High quality in modulation | Redundant acquisition | Whole slide imaging | ||
Separated detection | DHiLo17 | / | 0.44 | / | 53 | 5.0 | High robustness | Long post-processing time | Whole slide imaging | |
LiMo7 | / | 0.39 | 26 | 53 | 5.0 | High OS | Redundant acquisition | Whole brain imaging |
