Microscope Immersion Media: Oil vs Water vs Silicone

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What Are Microscope Objective Immersion Media?

Immersion media are liquids intentionally placed between the front lens of a high-magnification objective and the specimen’s cover glass (or directly onto the specimen in dipping configurations). Their central purpose is optical: by replacing the air gap with a material of higher refractive index, the objective can collect light over a larger cone angle. This increases the numerical aperture (NA), which directly improves both lateral resolution and light-gathering efficiency.

\"Principle — Principle of immersion microscopy. At high magnification power, light waves refract off the glass in the microscope slide and slip cover. Immersion oil has a high refractive index, minimizing this refraction allowing light to enter the objective in a straight line. This increases resolution of the specimen.
Artist: Thebiologyprimer

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In microscopy, numerical aperture is defined as NA = n sin θ, where n is the refractive index of the medium in contact with the objective’s front element and θ is half the angular aperture of the light cone accepted by the objective. Air has n ≈ 1.00; typical immersion oils are formulated around n ≈ 1.515 at visible wavelengths to match glass; water has n ≈ 1.333; glycerol is around n ≈ 1.47; and silicone immersion media are commonly around n ≈ 1.40–1.41. Using a medium with higher n raises the achievable NA for the same mechanical geometry, which improves resolving power.

Because resolution in widefield imaging scales with NA, immersion media are a critical accessory for micrographs that demand fine detail. They also influence spherical aberration, field flatness, depth of field, and reflective losses at interfaces. The choice among oil, water, glycerol, or silicone is not merely a preference—it is about physically matching the optics to the sample and mounting environment.

Oil immersion: optimized for fixed samples under standard cover glasses where matching the refractive index of glass reduces aberrations and surface reflections.

Water immersion: favored for live, aqueous specimens and for imaging deeper into water-based samples to reduce index mismatch.

Glycerol immersion: useful when mounting media or specimen refractive index lies between water and oil, often in cleared or preserved samples.

Silicone immersion: designed for live-cell and thicker, soft tissues whose refractive indices are closer to cytoplasm, with excellent focus stability and reduced evaporation.

Each of these choices interacts with cover glass thickness, objective design, and sample properties. Understanding how these variables connect will help you use objectives as they were intended, avoiding the pitfalls that degrade image quality.

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Optical Foundations: NA, Refractive Index, and Resolution

Microscope image quality rests on a few core optical relationships. Mastering these will clarify why immersion media matter so much and when they deliver real gains.

Numerical aperture and light collection

Numerical aperture is defined by NA = n sin θ, where n is the refractive index of the medium that light traverses as it enters the objective’s front lens, and θ is half the angular width of the light cone that can reach the lens. Higher NA increases both spatial resolution and photon throughput. For a given θ, increasing n via immersion raises NA directly. That is why most objectives that push beyond NA ≈ 0.95 are oil, glycerol, or water immersion types.

Because NA contains n, even a moderate half-angle can yield very high NA if the medium has a high refractive index. For example, if sin θ = 0.95 (a wide cone), then:

Air (n ≈ 1.00): NA ≈ 0.95

Immersion oil (n ≈ 1.515): NA ≈ 1.44

This simplified comparison shows why immersion can make such a difference to the finest details a microscope can resolve.

Resolution and its dependence on NA

In incoherent, widefield microscopy, two common expressions describe lateral resolution limits:

Abbe criterion: d ≈ λ / (2 NA)

Rayleigh criterion: d ≈ 0.61 λ / NA

These formulas relate the minimum resolvable distance d to the wavelength λ and the NA. Both predict improved resolution as NA increases. At λ = 550 nm and NA = 1.40, the Rayleigh distance is roughly d ≈ 0.61 × 550 / 1.40 ≈ 240 nm, while Abbe gives about 196 nm. The precise limit depends on imaging conditions, but the governing trend—higher NA yields finer detail—is unambiguous.

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Depth of field and trade-offs

As NA increases, depth of field (DOF) decreases. Approximate relations for widefield imaging scale as DOF ∝ λ / NA², ignoring detector and tube lens effects. Practically, higher-NA immersion objectives deliver razor-thin focal slices. That is often an advantage for rejecting out-of-focus haze but can make focusing and maintaining focus more demanding, especially in live specimens where the sample may drift. These trade-offs are an inherent part of the optical design, not a flaw.

If you need deeper focal penetration into thick, aqueous samples, water or silicone immersion can mitigate refractive index mismatch relative to oil, which, in turn, can reduce spherical aberration accumulation at depth.

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Oil, Water, Glycerol, and Silicone: A Practical Comparison

Four immersion families dominate modern light microscopy. They differ in refractive index, viscosity, volatility, environmental tolerance, cleaning difficulty, and their interactions with specimen and coverslip.

Oil immersion: the glass-matching standard

Modern synthetic immersion oils are formulated with refractive indices close to that of common optical glass and standard coverslips (n ≈ 1.515 at visible wavelengths). The primary benefit is index matching across the optical path—glass coverslip to oil to objective front lens—minimizing spherical aberration and surface reflections at those interfaces.

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When to consider oil immersion:

High-NA imaging (NA ≈ 1.25–1.40 and above) on fixed, thin samples mounted under a standard #1.5 cover glass.

Maximum lateral resolution and photon throughput near the coverslip plane.

Situations where long time-lapse evaporation must be minimized (oil does not evaporate quickly).

Typical cautions:

Oil and water do not mix. Switching media without thorough cleaning degrades image quality and can damage or contaminate surfaces.

Viscosity can trap bubbles if applied improperly; bubbles compromise resolution and contrast.

Some immersion oils can creep into mechanical interfaces if over-applied; careful technique and immediate cleanup help.

Reflection losses at a glass–air interface are about 4% per surface at normal incidence. With oil matching, reflectance can fall to a negligible level. Reduced reflections improve contrast and signal-to-noise at the detector.

Water immersion: the live-cell and aqueous sample ally

Water immersion objectives bring the immersion medium’s refractive index closer to that of aqueous mounting solutions and the cellular environment (n ≈ 1.33–1.34 for buffers and water). That reduces refractive index mismatch at the specimen and can improve imaging deeper into the sample compared with oil, which differs more in n.

Why choose water immersion:

Live, intact specimens in aqueous buffer where maintaining physiological conditions is key.

Samples with structures several micrometers or more below the coverslip, where water’s index is closer to the sample than oil.

Faster cleanup and less residue compared with oil, which is particularly convenient for multi-user environments.

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Considerations:

Water evaporates, especially in warm rooms, gradually changing the layer thickness and potentially shifting focus over time.

Some water-immersion objectives are designed for water-dipping (direct immersion into the aqueous sample without a coverslip), typically featuring longer working distances; others are designed for standard coverslips with a thin water layer between cover glass and front lens.

Maximum NA is typically lower than oil for a similar cone angle due to water’s lower n, though high-NA water objectives exist.

Glycerol immersion: a middle ground for cleared or preserved samples

Glycerol has a refractive index around 1.47 at visible wavelengths. That places it between water and oil, making it a useful compromise when the specimen or mounting medium has been adjusted toward higher n (e.g., some cleared tissues, high-glycerol mounting media, or matched buffers). A glycerol immersion objective can reduce spherical aberration compared with water or oil in such contexts.

When glycerol makes sense:

Specimens mounted in media with refractive indices in the mid-1.4s, where water and oil each produce notable mismatches.

Moderately high NA with less index mismatch than oil would present deeper into the sample.

Situations that call for slower evaporation than water but easier cleanup than some silicones.

Points to remember:

Glycerol is viscous and hygroscopic; its water content can change with ambient humidity over time, slightly shifting refractive index.

As with all immersion types, the objective should exactly match the intended immersion medium; using glycerol on a water-only or oil-only objective will not produce the designed performance.

Silicone immersion: stability and tissue-friendly refractive index

Silicone immersion media commonly have refractive indices around 1.40–1.41, not far from many biological tissues (n in the ~1.38–1.41 range for cytoplasm). This closer match helps limit index-induced spherical aberration when imaging tens of micrometers into soft, living samples. Silicone media are also known for low volatility, which helps maintain a stable layer over long time-lapse sessions.

Advantages of silicone immersion:

Improved focus stability in time-lapse due to low evaporation compared with water.

Refractive index closer to soft tissues, potentially reducing aberration with depth versus oil and sometimes even versus water.

Good wetting behavior on glass and objective fronts, helping maintain a continuous optical bridge.

Considerations:

Cleanup can be more involved compared with water or some oils; dedicated cleaning protocols recommended by the manufacturer are often necessary.

Silicone immersion objectives are specialized; using silicone on non-silicone objectives does not confer the same benefits and can compromise image quality.

In short, match the immersion medium to the objective and to the sample’s optical environment. If you adjust any one of these, re-evaluate the others to maintain performance. For a deeper dive into NA and resolution, revisit Optical Foundations.

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Cover Glass Thickness, #1.5H Tolerance, and Correction Collars

High-NA imaging is extremely sensitive to the geometry and optical properties of the cover glass. Most high-performance objectives are designed for a nominal cover glass thickness of about 0.17 mm (often labeled #1.5 or #1.5H). Deviations from this thickness, and variations in refractive index or flatness, introduce spherical aberration that softens detail and lowers contrast.

Standard vs high-precision covers

Common #1.5 coverslips are specified within a thickness range (often around 0.16–0.19 mm). High-precision (#1.5H) coverslips tighten that tolerance typically around 0.170 mm ± small variation (e.g., ± 0.005 mm). The tighter the tolerance, the closer the optical path length is to the objective’s design, which means sharper, more consistent images—especially at NA ≥ 1.3.

Even with the correct nominal thickness, manufacturing differences in glass and mounting media can leave residual aberrations. That is where correction collars help.

Correction collars: fine-tuning the optical path

Many high-NA objectives include a rotating correction collar. This ring slightly shifts internal lens spacing to compensate for small departures from the ideal optical path length caused by cover glass thickness, mounting medium, or temperature changes affecting refractive index. It is most beneficial when:

You must use coverslips that vary in thickness across slides.

You image at different wavelengths where dispersion might alter effective focus.

You work with aqueous samples prone to temperature-induced refractive index shifts.

Practically, you adjust the collar while inspecting fine, high-contrast features near the coverslip plane. The optimum setting maximizes sharpness and contrast across the field. If you move far below the coverslip, the original collar setting may no longer be optimal because spherical aberration grows with depth in mismatched media. Objectives designed for deep imaging with specific media (e.g., water or silicone) handle that better than collar tweaks alone.

For a refresher on why such small changes matter, see the definitions in Optical Foundations: NA, Refractive Index, and Resolution.

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Choosing the Right Immersion Medium for Your Sample

There is no universal best immersion medium; the right choice depends on the specimen, mounting conditions, and the objective’s optical design. The guiding idea is to minimize refractive index mismatches and geometric departures from the objective’s intended configuration.

Scenario 1: Thin, fixed specimens under standard coverslips

For histological sections, stained smears, thin polymer films, or microfabricated structures mounted under a #1.5 or #1.5H coverslip with a glass-like mounting medium, oil immersion objectives typically provide the highest NA and best lateral resolution near the coverslip. The oil’s refractive index closely matches both the coverslip and the objective’s front lens, reducing spherical aberration and reflection losses. This is the classic use case for oil immersion.

Scenario 2: Live, aqueous samples near the coverslip

Cells in buffer, plankton in water, or other living aqueous systems benefit from water immersion. The optical path more closely matches the specimen’s refractive index, reducing depth-dependent aberrations. A well-corrected water immersion objective can deliver crisp detail near the coverslip while avoiding the dramatic focus shifts that can occur if oil is used on thick, aqueous samples.

Scenario 3: Moderately thick specimens or media with elevated refractive index

When the mounting medium or sample index is significantly above water but below oil—such as high-glycerol buffers or partially cleared tissue—glycerol immersion often outperforms both water and oil. Its refractive index (~1.47) can provide a better overall match, improving contrast and resolution at depth compared with a large index mismatch.

Scenario 4: Live tissues and long time-lapse imaging

For thicker, soft tissues and extended time-lapse experiments where evaporation and thermal stability matter, silicone immersion has compelling benefits. Its refractive index near 1.40–1.41 is closer to cytoplasm and many soft tissues than oil, and it tends to maintain a stable layer over time. If your goal is to track fine dynamics over hours without frequent replenishment of the immersion layer, silicone immersion objectives can offer improved stability.

Scenario 5: Upright microscopes with dipping objectives

In aquatic ecology or electrophysiology setups, upright microscopes may use water-dipping objectives to reach directly into an aqueous bath without a coverslip. These lenses are engineered for long working distances and specific immersion geometries. Using them in the correct configuration is essential for maintaining designed NA and resolution. See also Working Distance, Sealing, and Mechanical Considerations for how geometry affects performance.

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Contrast, Aberrations, and Field Flatness with Different Media

Immersion media influence more than just NA. They modify reflections, aberrations, and even how evenly the field of view appears in focus.

Fresnel reflections and interface losses

Reflectance at a flat interface between two media at normal incidence is approximately R = ((n₁ − n₂) / (n₁ + n₂))². Consider these examples:

Glass–air (n ≈ 1.52 vs 1.00): R ≈ (0.52/2.52)² ≈ 0.0417, or ~4.2% per surface.

Glass–oil (n ≈ 1.52 vs ~1.515): The numerator is small, making R nearly negligible compared to glass–air.

Reducing these interface losses improves contrast because more signal reaches the detector and fewer internal reflections generate stray light. Oil immersion, designed to match glass, excels at reducing Fresnel losses at the coverslip–objective interface. Water and glycerol are not matched to glass as closely, but if they better match the specimen at depth, the overall result can still be higher contrast where it matters most.

Spherical aberration and refractive index mismatch

Spherical aberration arises when light rays passing through different zones of the pupil focus at different axial positions. In microscopy, a common source is refractive index mismatch between the immersion layer, coverslip, and specimen medium. The aberration tends to worsen with distance from the coverslip as light traverses mismatched materials. The net effect is a broadened point spread function (PSF), reduced peak intensity, and loss of high-spatial-frequency information.

Qualitatively, the wavefront error from mismatch grows with depth into the sample and with the magnitude of the index difference. Thus:

For oil on thick aqueous samples, mismatch can introduce notable spherical aberration a few micrometers beneath the coverslip.

For water or silicone on aqueous tissues, the mismatch is smaller, so depth performance can be better even if the nominal NA is slightly lower.

Correction collars allow some compensation near the coverslip but are not a cure-all for deep focus; choosing an immersion medium matched to the sample’s optical properties is often more effective than relying on collar adjustments alone. For the foundational math behind these trade-offs, see Optical Foundations.

Field flatness and chromatic effects

High-performance objectives strive for a flat field so that features at the same object-plane depth are equally in focus across the field of view. While field flatness is primarily a lens-design issue, immersion media that deviate from the design assumptions can exacerbate curvature of field and off-axis aberrations.

Dispersion (wavelength dependence of refractive index) also matters. Objectives labeled as apochromats are corrected across multiple wavelengths, but they are optimized for specific immersion conditions. If you switch immersion type or use a mounting medium with different dispersion, subtle lateral color shifts or chromatic blur can appear, especially off-axis. Keeping the immersion and mounting conditions close to the designer’s intent helps preserve apochromatic performance.

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Working Distance, Sealing, and Mechanical Considerations

Immersion media impose mechanical requirements. Tolerances, sealing, and working distance (WD) are tightly coupled to performance and usability.

Working distance vs NA

As NA increases, WD usually decreases. High-NA oil objectives often have very short working distances because capturing a wide light cone necessitates a large front lens and close approach to the coverslip. Water immersion objectives come in both short- and long-WD forms, especially for upright systems where water-dipping objectives must reach down into a bath. Always verify that the WD is sufficient for your sample thickness, chamber, and any protective windows in the optical path.

Sealing and compatibility

The front lens and barrel must resist the chosen immersion medium. Oils and silicones can creep; water can wick; glycerol can be hygroscopic. Objective housings and front lens cements are selected to withstand these effects for their intended media. Using a non-design medium risks chemical incompatibility, swelling of seals, or residue that is difficult to remove. When in doubt, consult the objective’s specifications for media compatibility.

Thermal stability and focus drift

Refractive indices change with temperature. Even small changes can shift the focus slightly because the optical path length through the immersion layer changes. Water layers thin by evaporation, causing additional drift in warm environments. Silicone immersion’s low volatility helps maintain layer thickness and focus position over long intervals, a key benefit in time-lapse imaging. If temperature control is part of your experiment, consider how your immersion medium responds to heating and cooling.

Mounting geometries and windows

Microfluidic devices, culture chambers, and environmental enclosures sometimes add glass or polymer windows between the objective and the sample. These modify the optical path. If the window material is not equivalent to a #1.5 glass coverslip of the expected thickness, you may see degraded performance, especially with oil immersion. Water or silicone immersion objectives designed for thicker windows or dipping modes can be more tolerant of such geometries. If you must image through additional windows, review the cover glass and correction collar considerations before choosing your immersion.

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Care, Cleaning, and Compatibility of Immersion Media

Maintaining pristine optics is essential for preserving resolution and contrast. Residues, films, and particulates on the front lens or coverslip scatter light and reduce image quality. Here are general, educational principles for caring for immersion optics. Always defer to the objective manufacturer’s guidance for specific solvents and methods.

General care principles

Use the intended medium. Oil for oil objectives, water for water objectives, glycerol for glycerol objectives, silicone for silicone objectives. Do not interchange.

Clean promptly. Remove immersion medium after use to avoid residue, dust capture, or creeping into lens edges.

Avoid abrasion. Use appropriate lens paper or swabs designed for optical surfaces; never use tissues that shed fibers or contain abrasive fillers.

Minimal solvent, minimal pressure. Apply small amounts to the paper or swab, not directly to the lens; use light, circular motions from center outward.

Media-specific considerations

Oil immersion residues can usually be removed with recommended lens-cleaning solutions or alcohols compatible with lens coatings, followed by a dry polish. Avoid aggressive solvents not specified by the manufacturer.

Water immersion leaves little residue; drying with lens paper is often sufficient, possibly followed by a gentle alcohol wipe if needed.

Glycerol is viscous and hygroscopic; gentle warm water followed by alcohol and a dry polish can help, within manufacturer guidelines.

Silicone immersion may require specialized, manufacturer-recommended cleaners to fully remove thin films. Do not assume oil cleaners will suffice.

Preventing cross-contamination

Dedicate slides and coverslips to a single medium when possible. Do not alternate water and oil on the same coverslip without cleaning.

Keep separate, clearly labeled applicators for each medium.

Inspect the front lens before and after each session. A hand loupe or the microscope itself (carefully focused on the front element) can reveal films or bubbles.

For how immersion media tie back to NA and resolution, revisit Optical Foundations. Selecting the correct medium is only part of the story; clean optics are equally vital to achieving the performance the physics promises.

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Worked Examples: NA, Resolution, and Reflectance

Back-of-the-envelope calculations help ground the concepts and set expectations. The following examples are generalized and intended for educational illustration.

Example 1: NA gains with immersion

Suppose an objective accepts a maximum half-angle such that sin θ = 0.95. Then:

Dry (air): NA = 1.00 × 0.95 = 0.95

Oil (n ≈ 1.515): NA ≈ 1.515 × 0.95 ≈ 1.44

Water (n ≈ 1.333): NA ≈ 1.333 × 0.95 ≈ 1.27

Glycerol (n ≈ 1.47): NA ≈ 1.47 × 0.95 ≈ 1.40

Silicone (n ≈ 1.41): NA ≈ 1.41 × 0.95 ≈ 1.34

Example 2: Resolution changes with NA

Using the Abbe expression d ≈ λ/(2NA) at λ = 550 nm:

NA = 0.95: d ≈ 550 / (2 × 0.95) ≈ 289 nm

NA = 1.27: d ≈ 550 / (2 × 1.27) ≈ 216 nm

NA = 1.40: d ≈ 550 / (2 × 1.40) ≈ 196 nm

NA = 1.44: d ≈ 550 / (2 × 1.44) ≈ 191 nm

The gains from 0.95 to 1.40 NA are substantial. Whether those theoretical gains translate to your sample depends on aberrations, scattering, and how well the immersion and cover glass match the objective design. The closer the path is to the designed conditions, the more faithfully the PSF approaches the diffraction limit.

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Example 3: Fresnel reflection reductions

At normal incidence:

Glass–air: R ≈ ((1.52 − 1.00) / (1.52 + 1.00))² ≈ (0.52 / 2.52)² ≈ 0.0417 or ~4.2% per surface.

Glass–oil: R ≈ ((1.52 − 1.515) / (1.52 + 1.515))² ≈ (0.005 / 3.035)², a fraction of a percent, effectively negligible in typical imaging contexts.

Reducing these reflections not only increases throughput but also suppresses ghosting and flare that wash out weak details. Oil immersion’s glass matching is particularly useful for brightfield and fluorescence where contrast is at a premium.

Example 4: Depth of field scaling

A simple scaling relation for widefield suggests DOF ∝ λ/NA². If you double NA, DOF drops by a factor of four (holding wavelength constant). That is one reason high-NA objectives produce crisp, thin optical sections but require careful focusing and stable specimens. The exact DOF depends on detection geometry, camera pixel size, and other factors, but the trend is universal.

Remember, this thinning of DOF is not a downside per se—when imaging flat specimens, it heightens contrast by rejecting out-of-focus background. In thick specimens, consider complementary strategies (appropriate immersion, careful mounting, and, if available, optical sectioning methods) to manage background light.

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Frequently Asked Questions

Can I use water on an oil-immersion objective (or vice versa)?

It is not recommended. Objectives are designed for a specific immersion medium; changing it alters the refractive index boundary conditions at the front lens, which changes the effective spherical aberration, focus, and overall performance. Using the wrong medium can also leave residues that are difficult to remove or risk damaging seals. Always use the immersion medium specified on the objective barrel, and see Care, Cleaning, and Compatibility for general maintenance principles.

Does silicone immersion affect fluorescence color or brightness?

Silicone immersion does not change emission wavelengths; it is not a filter. However, immersion media and mounting media can contribute background autofluorescence to varying degrees, and they can influence the point spread function via refractive index mismatch. Properly chosen silicone immersion formulated for fluorescence tends to have low autofluorescence, but always verify with your dyes and mounting conditions. For how immersion influences resolution and signal collection, revisit Optical Foundations and Contrast and Aberrations.

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Final Thoughts on Choosing the Right Immersion Medium

Selecting the right immersion medium is about more than achieving a high number on the objective barrel. It is about matching the optical path—cover glass thickness, refractive index of the specimen and mounting environment, and the objective’s internal design—to the realities of your sample. When those pieces align, the microscope delivers what physics allows: maximal resolution, improved contrast, and reproducible performance.

Use oil immersion when you need the highest NA at the coverslip for thin, fixed specimens under standard #1.5 or #1.5H glass. Prefer water or silicone immersion for live and aqueous samples, especially when imaging below the coverslip, to limit mismatch-induced spherical aberration. Glycerol immersion fills the middle ground when your mounting medium’s refractive index sits between water and oil. Do not overlook mechanical realities: working distance, stability, evaporation, and cleanup will shape your day-to-day experience as much as pure optical considerations.

To deepen your understanding, revisit the sections on NA and resolution, cover glass tolerance and correction collars, and contrast and aberrations. If you found this guide useful, consider subscribing to our newsletter to receive future articles on microscope fundamentals, accessories, and practical imaging tips.

Microscope Immersion Media: Oil vs Water vs Silicone

Table of Contents

What Are Microscope Objective Immersion Media?

Optical Foundations: NA, Refractive Index, and Resolution

Numerical aperture and light collection

Resolution and its dependence on NA

Depth of field and trade-offs

Oil, Water, Glycerol, and Silicone: A Practical Comparison

Oil immersion: the glass-matching standard

Water immersion: the live-cell and aqueous sample ally

Glycerol immersion: a middle ground for cleared or preserved samples

Silicone immersion: stability and tissue-friendly refractive index

Cover Glass Thickness, #1.5H Tolerance, and Correction Collars

Standard vs high-precision covers

Correction collars: fine-tuning the optical path

Choosing the Right Immersion Medium for Your Sample

Scenario 1: Thin, fixed specimens under standard coverslips

Scenario 2: Live, aqueous samples near the coverslip

Scenario 3: Moderately thick specimens or media with elevated refractive index

Scenario 4: Live tissues and long time-lapse imaging

Scenario 5: Upright microscopes with dipping objectives

Contrast, Aberrations, and Field Flatness with Different Media

Fresnel reflections and interface losses

Spherical aberration and refractive index mismatch

Field flatness and chromatic effects

Working Distance, Sealing, and Mechanical Considerations

Working distance vs NA

Sealing and compatibility

Thermal stability and focus drift

Mounting geometries and windows

Care, Cleaning, and Compatibility of Immersion Media

General care principles

Media-specific considerations

Preventing cross-contamination

Worked Examples: NA, Resolution, and Reflectance

Example 1: NA gains with immersion

Example 2: Resolution changes with NA

Example 3: Fresnel reflection reductions

Example 4: Depth of field scaling

Frequently Asked Questions

Can I use water on an oil-immersion objective (or vice versa)?

Does silicone immersion affect fluorescence color or brightness?

Final Thoughts on Choosing the Right Immersion Medium

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