Multimodal Microscope | SIMTRUM Photonics Store

Multi-module microscope optical path diagram

Front view of a multi-module microscope

Multimodal and multipattern   The Multimodal Switching Hub integrates three spatial excitation geometries (upright, side, inverted) and two light sources (laser/LED), supporting five operational modes: Single/dual-laser upright Single/dual-laser side Laser inverted LED upright LED side Mode switching takes <2 seconds via software or motorized sliders—no manual realignment needed. Factory-calibrated to sub-micron precision; optical loss <3%, pointing stability ≤ ±2 µm. This “plug-and-play” design dramatically improves experimental efficiency and data reproducibility across diverse samples—from solid films and PA nanocrystals to dynamic solutions—and from static imaging to ultrafast time-resolved studies.

Multimodal Operation Modes

Highly Integrated Multimodal Fluorescence Dynamics Testing System 

Sample Excitation Configuration

Spectral Resolution Acquisition Module	Spectral Resolution Acquisition Module Covering a broad spectral range from 200 to 1700 nm, the module achieves a spectral resolution of better than 0.1 nm in the visible region, enabling precise resolution of fine spectral features such as multi-peak structures, Stokes shifts, and energy-level splitting. It employs a high-transmission focusing optical path combined with low-loss fiber coupling technology to ensure efficient signal delivery to the spectral analysis unit. The accompanying intelligent software automatically performs background subtraction, peak identification, multi-peak fitting, intensity normalization, and component deconvolution, and can generate professional reports—complete with spectral curves and key parameters—with a single click. This fully supports applications ranging from basic characterization to in-depth mechanistic studies.
Project	Parameter
Range	200–1700 nm (UV to NIR-II)
Resolution	≤ 0.1 nm（@400 nm），≤ 0.5 nm（@1000 nm）
Detectors	Back-thinned CCD (200–1100 nm) + InGaAs array (900–1700 nm)
Dynamic Range	>10⁶:1
Modes	Continuous/step scanning, time-series
Calibration	Built-in Hg/Ar lamp for auto wavelength correction
Features	Auto nonlinear index fitting (I ∝ Pⁿ, n >500) Multi-peak Gaussian/Lorentzian fitting Stokes shift calculation Automatic identification of Tm³⁺/Er³⁺ peaks (e.g., ¹G₄→³H₆ @475 nm)

Time-resolved detection module	Time-resolved detection module Based on Time-Correlated Single Photon Counting (TCSPC) technology, the system achieves a time resolution better than 50 ps, enabling precise measurement of dynamic processes across the full timescale—from nanosecond fluorescence decays to second-scale afterglow emissions. It supports simultaneous acquisition of multidimensional parameters such as rise time, decay lifetime, and delayed luminescence intensity, offering comprehensive insights into excited-state energy relaxation, charge transfer, or triplet-state evolution mechanisms. Equipped with highly sensitive detectors—including avalanche photodiodes (APDs), single-photon counting modules (SPCMs), or high-speed photomultiplier tubes (PMTs)—the system delivers high signal-to-noise ratio lifetime curves even under extremely low-concentration or weak-emission conditions. It serves as an essential tool for dynamic studies of advanced luminescent materials such as TADF emitters, phosphors, upconversion systems, and quantum dots.
Project	Parameter
Time Resolution	<30 ps
Measurement Range	100 ps – 10 s (auto-ranging)
Excitation Sync	Ps-pulsed laser (1 MHz – 80 MHz rep rate)
Detectors	MCP-PMT (200–900 nm) or NIR SPAD (900–1700 nm)
Fitting Models	Mono/bi/tri-exponential, stretched exponential, IRF deconvolution
Features	PA dynamics package: decouples ³F₄ (cycling) and ¹G₄/³H₄ (emission) lifetimes Compatible with Nature 2025 data format (e.g., ³F₄ ≈ 95 µs, ³H₄ ≈ 26 µs) Auto-output: τ₁, τ₂, A₁, A₂, χ² Rise time (Tᵣᵢₛₑ) and avalanche buildup kinetics

Photon Avalanche (PA) Materials

Enables full-parameter characterization of ultra-nonlinear emitters (e.g., NaLuF₄:Tm³⁺ nanocrystals), including:

• Nonlinear exponent (n > 500)
• Avalanche threshold (～16 kW/cm²)
• Single-particle super-resolution imaging (<50 nm)
• Supports development and mechanistic studies of next-generation PA probes (Nature 2025).

 Super-Resolution Optical Imaging

Leverages extreme nonlinearity of PA materials to achieve <50 nm resolution using a single continuous-wave laser—eliminating the need for complex STED or RESOLFT setups. Ideal for low-phototoxicity, long-term live-cell imaging.

Ten-thousand-particle Monte Carlo simulations validate the universality of PA-based super-resolution.

Fluorescent Material Optimization & Screening

Combines spectral and lifetime data to rapidly evaluate:

• Molecular design
• Doping ratios
• Core-shell engineering
• Synthesis protocols
• Accelerates iteration of high-performance upconversion, TADF, and long-afterglow materials.

XRD Rietveld refinement reveals lattice contraction (Δa ≈ −0.5%) due to Lu³⁺ substitution.

Fluorescent Probe Development

Validates targeting, sensitivity, and biocompatibility of smart probes for pH, ions, ROS, etc.—especially NIR-II PA probes for dynamic monitoring.

Surface ligand engineering significantly influences the PA emission intensity and stability.

Photophysical Dynamics Studies

Precisely resolves transient behaviors: fluorescence lifetime, rise/decay times, delayed emission. Reveals microscopic mechanisms like excited-state relaxation, cross-relaxation (CR), and energy migration—providing quantitative inputs for multi-level systems (e.g., Tm³⁺/Er³⁺).

Host engineering modulates the Tm³⁺ energy level lifetimes to optimize PA dynamics.

Nanomaterial Spectral Analysis

Performs single-particle analysis of:

• Emission peak position
• FWHM
• Quantum yield
• Blinking statistics
• Enables quality control and batch consistency for quantum dots, perovskites, MOFs/COFs, and PA nanocrystals.

XPS reveals enhanced local crystal field, leading to improved luminescence efficiency.

Interdisciplinary Research & Industrial Testing

Applications span:

• Life sciences
• Optoelectronic devices
• Anti-counterfeiting tags
• IR displays
• Chemical sensing
• High-throughput material screening
• Bridges lab-scale discovery to industrial production.
• PA materials in anti-counterfeiting, display, and sensing,Multiplexed bio-detection via multi-channel PA imaging

Validation of the System’s Multimodal Operating Modes

This example demonstrates the system’s five core operating modes through specific experiments. In each mode, digital image acquisition, spectrally resolved acquisition, and time-resolved acquisition are successfully implemented, thereby verifying the system’s high level of integration and flexible reconfigurability.

Overall Architecture

Figure 1: Schematic diagram of the overall optical layout of the system, illustrating the integrated configuration of the laser/LED excitation sources, the multimodal functional module (4), the sample excitation modules (5, 6, 7), and the three acquisition modules (8, 9, 10).

Upright Single/Dual-Beam Laser Excitation Mode

Optical Layout Schematic: Figures 2(a–c)

Functional Description:

• Figure 2(a): Illustrates the digital image acquisition path. The excitation light is focused onto the sample through the objective lens, and the emitted fluorescence is collected by the same objective lens and directed to the imaging sensor.
• Figure 2(b): Depicts the spectrally resolved acquisition path. The emitted fluorescence is purified by a filter and then directed into a spectrometer for wavelength-resolved analysis.
• Figure 2(c): Shows the time-resolved acquisition path. The emitted fluorescence, after spectral purification by a filter, is routed to a time-resolved photon detection unit to measure the luminescence rise time and lifetime.
• Figure 2(d): Presents a schematic of the light-path steering components within the internal submodules of the multimodal functional configuration module, ensuring proper routing of both excitation light and emission signals to their respective modules.

Lateral Single/Dual-Beam Laser Excitation Mode

Optical Layout Schematic: Figures 3(a–c)

Functional Description:

• Figure 3(a): Illustrates the digital image acquisition path. The excitation light is incident on the sample from the side, and the emitted fluorescence travels back along the same path to the imaging sensor.
• Figure 3(b): Depicts the spectrally resolved acquisition path. The emitted fluorescence is purified by a filter and then directed into a spectrometer for wavelength-resolved analysis.
• Figure 3(c): Shows the time-resolved acquisition path. The emitted fluorescence, after spectral purification by a filter, is routed to a time-resolved photon detection unit to measure the luminescence rise time and lifetime.
• Figure 3(d): Presents a schematic of the light-path steering components within the internal submodules of the multimodal functional configuration module, ensuring proper routing of both excitation light and emission signals to their respective modules.

Upright Single-Beam LED Excitation Mode

Optical Layout Schematic: Figures 4(a–c)

Functional Description:

• Figure 4(a): Illustrates the digital image acquisition path. Excitation light emitted by the LED source is focused onto the sample through the objective lens, and the resulting fluorescence is collected by the same objective lens and directed to the imaging sensor.
• Figure 4(b): Depicts the spectrally resolved acquisition path. The emitted fluorescence is purified by a filter and then directed into a spectrometer for wavelength-resolved analysis.
• Figure 4(c): Shows the time-resolved acquisition path. The emitted fluorescence, after spectral purification by a filter, is routed to a time-resolved photon detection unit to measure the luminescence rise time and lifetime.
• Figure 4(d): Presents a schematic of the light-path steering components within the internal submodules of the multimodal functional configuration module, ensuring proper routing of both excitation light and emission signals to their respective modules.

Lateral Single-Beam LED Excitation Mode

Optical Layout Schematic: Figures 5(a–c)

Functional Description:

• Figure 5(a): Illustrates the digital image acquisition path. Excitation light emitted by the LED source is incident on the sample from the side, and the resulting fluorescence travels back along the same path to the imaging sensor.
• Figure 5(b): Depicts the spectrally resolved acquisition path. The emitted fluorescence is purified by a filter and then directed into a spectrometer for wavelength-resolved analysis.
• Figure 5(c): Shows the time-resolved acquisition path. The emitted fluorescence, after spectral purification by a filter, is routed to a time-resolved photon detection unit to measure the luminescence rise time and lifetime.
• Figure 5(d): Presents a schematic of the light-path steering components within the internal submodules of the multimodal functional configuration module, ensuring proper routing of both excitation light and emission signals to their respective modules.

Inverted Single-Beam Laser Excitation Mode

Optical Layout Schematic: Figures 6(a–c)

Functional Description:

• Figure 6(a): Illustrates the digital image acquisition path. The excitation light is introduced from beneath the sample, and the emitted fluorescence is collected by an objective lens positioned above the sample and directed to the imaging sensor.
• Figure 6(b): Depicts the spectrally resolved acquisition path. The emitted fluorescence is purified by a filter and then directed into a spectrometer for wavelength-resolved analysis.
• Figure 6(c): Shows the time-resolved acquisition path. The emitted fluorescence, after spectral purification by a filter, is routed to a time-resolved photon detection unit to measure the luminescence rise time and lifetime.
• Figure 6(d): Presents a schematic of the light-path steering components within the internal submodules of the multimodal functional configuration module, ensuring proper routing of both excitation light and emission signals to their respective modules.

System Architecture and Principle

The system is based on an advanced multimodal optical design that supports multiple excitation modes and signal acquisition pathways. The schematic below illustrates the optical layout, with key modules including:

• Dual-laser input: Independent power control for 808 nm and 980 nm lasers
• Multi-channel signal acquisition: Simultaneous imaging, spectral, and time-resolved detection

Highly Integrated Multimodal Optical Architecture

Experimental Setup and Results

To validate the system’s dual-wavelength co-excitation capability, we selected two representative rare-earth-doped nanomaterials:

• NaYF₄:Tm/Nd – emits strong blue light (～475 nm) under 808 nm excitation
• NaYF₄:Yb/Er – emits green (～545 nm) and red (～655 nm) light under 980 nm excitation

The emission spectra under different excitation conditions are shown in the figure below:

• 808 nm excitation only: Strong blue emission from Tm³⁺ at ～475 nm
• 980 nm excitation only: Green (545 nm) and red (655 nm) emissions from Er³⁺
• 808 nm + 980 nm co-excitation: Simultaneous appearance of characteristic blue, green, and red emission peaks, with no spectral crosstalk and high signal-to-noise ratio

Typical Application Scenarios

• Characterization of Hybrid Systems Combining Photon Avalanche Materials and Conventional Upconversion Materials:Dual-wavelength excitation enables simultaneous activation of distinct energy transfer pathways, revealing the complex optical mechanisms within the material.
• Investigation of Competing Multi-Sensitization Pathways:Studying how different excitation wavelengths influence luminescent behavior allows optimization of excitation conditions to achieve optimal performance.
• Excitation Parameter Optimization Prior to Super-Resolution Imaging:Before performing super-resolution imaging, the system enables precise tuning of excitation parameters to ensure optimal fluorescence response from the sample.

To help users better understand why specific excitation wavelengths are required, the following diagram illustrates the photon avalanche energy-level mechanism in Tm³⁺-doped systems:

The system not only offers robust dual-wavelength co-excitation capability but also features high flexibility and user-friendliness, making it well-suited for research and development of advanced luminescent materials. Both academic laboratories and industrial R&D departments can benefit significantly from this platform, accelerating scientific discovery and technological translation.

Fluorescence Imaging and Spatial Localization Validation of Single Microrods

The system supports high-resolution brightfield–fluorescence dual-modal imaging, enabling precise spatial localization and visualization of luminescent properties of individual rare-earth-doped micro- or nanostructures (e.g., NaYF₄:Yb/Er microrods). This capability provides essential support for super-resolution microscopy, single-particle tracking, and micro-region spectral analysis.

Scientific Context and Link to Published Work

In Nature 2025 (DOI: 10.1038/s41586-025-09164-y), the research team achieved ～90 nm resolution in super-resolution imaging by exploiting the nonlinear photon avalanche response of Tm³⁺-doped nanocrystals (Main Text, Figure 4).

A critical prerequisite for this achievement was the precise localization of individual emitting particles and exact spatial correspondence between fluorescence signals and brightfield morphology.

This system serves as the core instrument that fulfills this prerequisite.

Experimental Setup and Results

NaYF₄:Yb/Er(18/2%) microrod samples were dispersed on a glass slide and mounted on an upright motorized 3D sample stage. The system was switched to the upright single-beam laser excitation mode, using a 980 nm laser for excitation. A scientific-grade sCMOS camera simultaneously acquired:

• Brightfield image (laser off)
• Fluorescence image (laser on, with a 500–700 nm bandpass filter)

(a): Brightfield image clearly resolves the microrod morphology (approximately 5 μm in length and 1 μm in width).

(b): Under 980 nm excitation, the same region exhibits strong green fluorescence from Er³⁺ (⁴S₃/₂ → ⁴I₁₅/₂).

Key validation: All fluorescent particles align precisely with their corresponding brightfield structures—no spatial offset or imaging artifacts observed.

Direct Integration with Super-Resolution Research

*The imaging capabilities of this system have successfully supported key experiments in the publication

Detailed Technical Advantages

Typical Application Scenarios

• Pre-processing for super-resolution microscopy: Screening isolated, high-brightness particles for subsequent photon avalanche (PA) or STED imaging.
• Characterization of micro/nanophotonic devices: Locating emission hotspots in individual microrods or nanowires.
• Biological labeling validation: Confirming successful binding of probes to single cells or subcellular structures.
• Educational demonstrations: Visually illustrating the integrated concept of “morphology + function” correlative characterization.

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