Buyers' Guides

Comparing Imaging Modes in Benchtop Microscopes: Widefield, Confocal, Super-Resolution and Transmitted Light Options

Author: Judith Beer

Published: 01 Jul 2026 · Last updated: 06 Aug 2026

The imaging mode describes the method a microscope uses to generate an image, defined by its illumination approach, detection strategy, and contrast mechanism. As each imaging mode differs in contrast, resolution, imaging depth, sensitivity, and speed, certain modes are better suited to specific sample types or experimental questions than others.

Benchtop fluorescence microscopes can vary widely in their capabilities and in the imaging modes they support. This overview explains the most common imaging modes available in benchtop systems and highlights the key factors to consider when evaluating a microscope.

Choosing a Benchtop Fluorescence Microscope – Which imaging modes should I consider?

Understanding which imaging modes are supported by a microscope is essential to ensure that the system can effectively address your biological or analytical goals. It is also important to note that imaging modes can vary significantly in technical complexity and cost, which can have a major impact on the overall price of the instrument.

Deconvolution image neuronal network drosophila

Transmitted Light Imaging

Generates label-free contrast by collecting light passing through the sample. Commonly integrated into benchtop fluorescence microscopes for quick sample assessment, live-cell monitoring, focusing, and added structural context. Contrast performance and costs depend strongly on the specific technique.

Transmitted-light technique	Description
Brightfield (BF)	Simplest and most cost-efficient option. Suits thicker or stained samples (e.g. histology sections).
Phase Contrast	Enhances contrast in thin samples using phase plate rings in the optical path. Suitable for imaging thin and even transparent samples.
Darkfield (DF)	Highlights high-transparency, low-contrast specimens. Typically limited to lower-magnification objectives and provides less structural detail than Phase Contrast, DIC or DPC.
Differential Interference Contrast (DIC)	Delivers high contrast and high resolution. Requires specialised optical components and is incompatible with plastic dishes.
Differential Phase Contrast (DPC)	Cost-effective and user-friendly alternative to DIC. Provides high-contrast, high-resolution imaging of thin samples without specialised optics. Compatible with plastic dishes.

Widefield Fluorescence Imaging

Illuminates the entire field of view simultaneously and collects both in-focus and out-of-focus emitted light. This is a fast, gentle, and cost-efficient imaging mode well-suited for routine fluorescence imaging and thin samples. However, it has limited depth resolution due to out-of-focus signal and does not suit thick and strongly scattering specimens.

Confocal Fluorescence Imaging

Uses optical sectioning to improve contrast and resolution and allows for high-quality 3D-imaging. It requires laser-based illumination and pinholes to reject out-of-focus light.

Confocal implementation	Description
Point-scanning confocal microscopy	Performs sequential scanning of the sample one point at a time. Offers high spatial resolution but has lower acquisition speeds and signal sensitivity. High phototoxicity and slow acquisition speeds can limit suitability for live-cell studies. Uses PMTs or GaAsP detectors with quantum efficiencies up to 45%.
Multi-point (spinning-disk) confocal microscopy	Uses multiple pinholes simultaneously to increase imaging speed and throughput. Reduces phototoxicity and signal loss, suited for sensitive specimens and live-cell imaging. Employs sensitive camera-based detectors with higher quantum efficiency >80%. Some systems incorporate dual microlens technology for improved light throughput, lower system background, higher signal to noise image, less sample phototoxicity and increased capacity for sample penetration and imaging at depth. Learn more about point scanning and spinning disk confocal microscopes.

Super-Resolution Imaging

Techniques that exceed the diffraction limit of conventional light microscopy (~200 nm, wavelength-dependent), enabling visualisation of structures down to ~10–150 nm, depending on the method. Hardware-based implementations are often complex and cost intensive. Not common in benchtop systems, but in some cases available.

Super-resolution technique	Description
SIM (Structured Illumination Microscopy)	Uses patterned light projected onto the sample and computational reconstruction. Resolution ~100-120 nm. Requires specific hardware, uniform samples and stable dyes. Compatible with fixed and live samples. Expensive, but lower cost than STED.
STED (Stimulated Emission Depletion Microscopy)	Uses a focused depletion laser to suppress fluorescence surrounding the focal spot. Resolution ~30–50 nm. Requires complex and expensive hardware, high laser power, special sample preparation, and specialised, highly stable dyes. Typically not suited for live-cell imaging.
SMLM (Single-Molecule Localisation Microscopy techniques, e.g. STORM)	Software-driven approach requiring high level of expertise. Relies on very high laser powers and acquisition of thousands of imaging frames to pinpoint molecule localisation. Achieves resolutions down to 10 nm. Generally unsuitable for live-cell imaging.
SRRF-Stream (Super-Resolution Radial Fluctuations)	Software-based super-resolution technique using acquisition of 10–100 frames per field of view. Compatible with standard fluorophores, does not require special sample preparation. Resolution down to ~140 nm. Suitable for live-cell imaging and fixed thick samples.

Multimodality

Multiple imaging modes available in a single benchtop platform enable flexibility across diverse samples and experimental workflows. This is typically associated with higher system costs. Modular systems may allow staged upgrades to scale functionality and investment over time.

Representative images of imaging modalities

Fig. 1 — Representative images of imaging modalities available in BC43. Top images: (left) Confocal image of Drosophila embryos stained with a neuronal marker. (middle) Widefield & widefield-Clearview (deconvolution), mammalian cells showing actin, DNA, mitochondria, and microtubules. Images were deconvolved. (right) DPC and DPC image overlapped with a confocal image. Bottom images: The same cell was imaged using three different imaging modalities: widefield (left), confocal (centre), and super resolution (right). The increase in detail and resolution is clearly observed when different imaging modalities are used. Image credits: Alvaro Tavares and Marco Campinho, Universidade do Algarve, and Claudia Florindo, Oxford Instruments. Confocal (Drosophila) – CF & AT; DPC & Confocal – CF & MC; all others – CF.

Exploring Benchtop Fluorescence Microscopes?

Learn more about the key technologies, components and practical considerations involved in selecting a benchtop fluorescence microscope:

Microscope Chassis

Sample Stage

Illumination

Filters and Mirrors Filters & Mirrors

Objectives

Detectors

Vibration and Environmental Control Vibration & Environmental Control

Software

Quality Control, Upgradability & Service

BC43 – Exceptional Performance, Certified Quality, High Productivity

BC43 is a multimodal imaging system supporting transmitted light (brightfield and DPC), widefield, confocal imaging, and software-based super resolution (SRRF-Stream). The BC43 entry model featuring widefield imaging can be upgraded in the field to confocal with minimal system downtime, optional super-resolution can be added flexibly at any model (Widefield or confocal).

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