Revvity IVIS Spectrum 2 Small Animal In Vivo Optical Imaging System

Overview

The Revvity IVIS Spectrum 2 is a high-performance, modular small animal in vivo optical imaging system engineered for quantitative bioluminescence and fluorescence detection in murine models. Based on cooled, back-illuminated CCD technology enhanced with proprietary eXcelon® anti-reflective coating, the system achieves exceptional quantum efficiency (>95% at 600–800 nm) and ultra-low dark current (<0.001 e⁻/pixel/sec), enabling detection of weak photon signals from deep-tissue reporter expression or low-abundance fluorescent probes. Unlike conventional widefield optical imagers, the IVIS Spectrum 2 integrates spectral unmixing, automated focus calibration, and motorized FOV adjustment to support both 2D planar imaging and 3D tomographic reconstruction (BLT/FLT). Its optical architecture complies with ISO 17025 traceable calibration protocols and supports longitudinal, non-invasive monitoring of biological processes—including tumor progression, stem cell trafficking, gene therapy expression, pharmacokinetics, and immunotherapeutic response—under controlled vivarium conditions.

Key Features

• Proprietary eXcelon®-coated, thermoelectrically cooled CCD sensor with 1024 × 1024 pixel resolution and <1 e⁻ RMS read noise
• Motorized, dual-lens optical path supporting 10 discrete FOVs (3.9 × 3.9 cm to 22.5 × 22.5 cm) with auto-focus and auto-exposure calibration
• Spectral unmixing engine capable of resolving ≥4 spectrally distinct fluorophores (e.g., Cy5.5, IRDye800CW, Alexa Fluor 750) while suppressing endogenous autofluorescence across 400–900 nm
• Integrated 3D reconstruction module using diffuse optical tomography (DOT) algorithms validated against NIST-traceable phantom standards
• High-throughput imaging mode with programmable mouse positioning stage and smart tray recognition—supports concurrent imaging of up to 10 anesthetized mice with individual ROI annotation
• Modular docking interface compliant with ASTM F2965-14 for seamless integration with micro-CT (e.g., PerkinElmer Quantum GX or Bruker Skyscan systems) for anatomical-functional co-registration

Sample Compatibility & Compliance

The IVIS Spectrum 2 is optimized for use with immunocompromised and transgenic mouse strains (e.g., NSG, BALB/c nude, C57BL/6) under IACUC-approved protocols. It accommodates standard rodent anesthesia delivery systems (isoflurane vaporizer + nose cone manifold) and supports temperature-controlled stages (30–37 °C) to maintain physiological stability during acquisition. All optical filters meet ISO 10110-7 surface quality specifications; excitation/emission filter sets are certified for >OD6 blocking and <±2 nm center wavelength tolerance. The system adheres to FDA 21 CFR Part 11 requirements for electronic records and signatures when deployed with validated Living Image™ software v8.0 or later, including full audit trail logging, user role-based access control, and electronic signature workflows for GLP/GMP-compliant preclinical studies.

Software & Data Management

Controlled exclusively via Living Image™ software (v8.0+), the IVIS Spectrum 2 provides a validated, FDA 21 CFR Part 11-compliant environment for acquisition, quantification, and reporting. The software features wizard-driven experimental setup, batch processing pipelines for multi-session longitudinal analysis, and built-in statistical modules (ANOVA, t-test, dose-response curve fitting). Raw photon count data are stored in vendor-neutral HDF5 format with embedded metadata (animal ID, anesthesia duration, exposure time, filter set, binning factor). Export options include DICOM-SR (Supplement 122), MIAME-compliant MAGE-TAB, and CSV/TXT for integration into downstream platforms such as MATLAB, Python (via PyIVIS), or commercial PK/PD modeling tools (e.g., Phoenix WinNonlin). Audit trails record every parameter change, image export, and ROI modification with timestamp, operator ID, and reason-for-change field.

Applications

• Longitudinal monitoring of orthotopic and metastatic tumor burden using luciferase-expressing xenografts
• Quantitative assessment of protease activity (e.g., MMP-2/9) via activatable near-infrared probes
• Tracking of adoptively transferred immune cells labeled with NIR fluorophores or luciferase reporters
• Pharmacodynamic evaluation of targeted therapeutics through real-time reporter gene induction (e.g., NF-κB, HIF-1α)
• Multi-parametric validation of CRISPR/Cas9 editing efficiency in vivo using dual-reporter constructs
• Correlative imaging studies combining functional optical data with micro-CT-derived bone volume, lung density, or vascular architecture

FAQ

What is the minimum detectable photon flux for bioluminescence imaging?
The system achieves a theoretical limit of detection of ≤100 photons/sec/cm²/sr under optimal conditions (f/1.2 lens, 5-min exposure, −90°C sensor), validated using NIST-traceable calibrated light sources.
Can the IVIS Spectrum 2 perform fluorescence lifetime imaging (FLIM)?
No—this system is designed for intensity-based spectral unmixing and does not incorporate time-correlated single-photon counting (TCSPC) hardware required for FLIM.
Is remote operation supported for BSL-2 or ABSL-2 facilities?
Yes—Living Image™ supports secure client-server deployment over VLAN-separated networks with TLS 1.2 encryption and optional LDAP/Active Directory integration.
How is calibration maintained across imaging sessions?
Daily automated flat-field correction and weekly photometric calibration using integrated LED reference source ensure inter-session reproducibility (CV <3.5% over 30 days, per Revvity IQ/OQ documentation).
Does the system support intravital microscopy integration?
Not natively—the IVIS Spectrum 2 is a macroscopic whole-body imager; however, co-registered datasets can be spatially aligned with confocal or two-photon histology using landmark-based affine transformation in Living Image™.