Comparison of immersion objective types

Water immersion objective features

  • Best for observing cultured cells and other specimens in culture solution because there is little spherical aberration as the refractive index is close to that of cultured cells and culture solution
  • Best for in vivo observations and live tissue fragments because it is highly biocompatible as the refractive index is close to that of biological fluids
  • Also effective with water dispensers when it is necessary to supply immersion fluid, such as for observations of multiple points in multiple wells

Silicone immersion objective features

  • Little spherical aberration when making deep observations because the refractive index is close to that of many 3D cell cultures (spheroid, organoid, etc.) and organs with high refractive index (liver, etc.)
  • Best for long time-lapse observations because oil stable and resistant to drying even at 37°C.

Oil used for silicone immersion objective: Silicone immersion oil 30cc

Oil immersion objective features

  • Appropriate for observations that require high resolution because it has the highest NA of all immersion objective lenses
  • Best for observations of sample surfaces (vicinity of cover glass) because its refractive index is close to that of cover glass (ne=1.53)
  • Best for observations of anchored specimens that use a mounting medium because the refractive index is close to that of many mounting mediums

Glycerin immersion (cleared specimen) objective lens features

  • Possible to make deep observations of cleared specimens because glycerin has a refractive index close to many common clearing agents
  • Can use with a wide range of clearing agents
  • Can optimize spherical aberration correction to match the refractive index of the clearing agent using the correction collar

Examples of clearing agents appropriate for glycerin immersion (for clearing) objective lens:

Sca/e(n=1.38)*, CUBIC2(n=1.48), LUCID(n=1.47), FocusClear(n=1.45), CRARITY(n=1.45), SeeDB(n=1.48)

*Refractive index depends on the organism

Comparison of immersion objective features

Immersion medium
(possible magnification range)
Water (20X-60X) Silicone (25X-100X) Oil (40X-100X) Glycerin (10X-20X)
Immersion liquid refractive index Water (ne=1.33) Silicone (ne=1.41) Oil (ne=1.52) Clearing agent (ne=1.33~1.51:10X, ne=1.44~1.50: 20X), Glycerin (ne=1.45)
Appropriate live samples for observation Cultured cells
Animal tissue and organs (mouse brain, etc.) with low refractive index
Observations that must be biocompatible (in vivo observations, etc.)
Observations that require multi-point observations of multiple wells (use of water dispensers)
3D cell cultures (spheroid, organoid, etc.)
Animal tissue and organs with high refractive index (liver, etc.)
Long time-lapse observations
Cultured cells and tissue fragments that require high resolution (sample surface)
Appropriate anchored samples for observation Anchored cultured cells
Anchored organ fragments with low refractive index (mouse brain, etc.)
Anchored fragments, such as organs with high refractive index (liver, etc.) Cultured cells (with mounting medium)
Tissue fragment (with mounting medium)
Cleared samples (spheroid, organoid, organs, etc.)