Overview

TruSpectral High-Sensitivity Multichannel Imaging

Using proprietary spectral detection technology, the FV3000 confocal microscope’s TruSpectral detectors combine high sensitivity with spectral flexibility to detect even the dimmest fluorophores.

  • Up to 3X more light transmission vs. traditional spectral detection technology
  • Independently adjustable channels to optimize signal detection for each individual fluorophore
  • Lambda scanning mode enables accurate spectral unmixing of complex overlapping fluorescent signals
  • Variable barrier filter mode provides simultaneous four-channel image acquisition, with up to sixteen channels in virtual channel mode

Macro-to-Micro Imaging and Super Resolution Microscopy

The FV3000 microscope’s macro-to-micro workflow provides a roadmap for data acquisition, enabling you to see data in context and easily locate regions of interest for higher resolution imaging.

  • Use a low-magnification 1.25X or 2X objective to quickly capture a large field of view (FOV) map of whole specimens
  • Identify regions of interest on the Overlay Map, then switch to a higher magnification objective for high-resolution confocal imaging down to 120 nm with Olympus Super Resolution technology (FV-OSR)
  • Finalize your acquisition and get publication-ready microscope images with TruSight image processing

Mouse brain hemisection embedded for expansion microscopy (pre-expansion), labeled with secondary antibodies against GFP (Alexa Fluor 488, green), SV2 (Alexa Fluor 565, red), Homer (Alexa Fluor 647, blue).
Sample courtesy of Dr. Ed Boyden and Dr. Fei Chen, MIT.

Dendrite (anti-GFP Alexa Fluor 488, green) and synaptic marker (SV2, Alexa Fluor 565, red). Olympus Super Resolution image processed with cellSens advanced constrained iterative deconvolution. Average full width half maximum measurements ~135 nm. Image acquired with 100X 1.35 NA silicone objective.
Sample courtesy of Dr. Ed Boyden and Dr. Fei Chen, MIT.

Hybrid Scanning for High-Speed Imaging and Increased Productivity

The FV3000 Hybrid Scanner provides two scanners in one for enhanced confocal imaging capabilities.

  • The FV3000RS hybrid scan unit uses a galvanometer scanner for precision scanning as well as a resonant scanner, ideal for high-speed imaging of live physiological events
  • Capture video-rate images with a large FOV using the resonant scanner, featuring speeds from 30 frames per second (fps) at FN 18 all the way up to 438 fps using clip scanning
  • Use the resonant scanner to observe fast phenomena, such as a beating heart, blood flow, or calcium ion (Ca2+) dynamics inside cells
  • Switch between the galvanometer scanner and resonant scanner with the click of a button

Accurate Time-Lapse Imaging

Time-lapse experiments require consistent focus and low phototoxicity to the sample.

  • Olympus’ TruFocus unit helps maintain focus during live cell imaging despite changes in temperature or added reagents
  • The FV3000 microscope’s high-sensitivity detector requires significantly less laser power while the resonant scanner reduces laser illumination time, lowering phototoxicity for more physiologically accurate confocal imaging data

Deep Tissue Observation with Silicone Objectives

The refractive index of silicone oil is close to that of living tissue, enabling high-resolution observation deep inside tissue with minimal spherical aberration.

  • Refractive index match delivers an ideal focal volume, resulting in perfect volume reconstruction and enabling high-resolution confocal imaging of large living organisms
  • Long working distances enable detailed microscopic imaging at depth
  • See data unfold in real-time and easily observe structures with 3D reconstruction software

Applied Technology

TruSpectral Detection

Groundbreaking TruSpectral detection technology offers outstanding results compared to previous generations of spectral detection units. Featured on every channel in the FV3000 microscope, TruSpectral detection technology combines the flexibility of a spectral detector with the sensitivity of a filter-based detector.

How TruSpectral Detection Technology Works

Based on patented volume phase hologram (VPH) transmission for high efficiency, TruSpectal detection uses an adjustable slit to select the detection wavelengths of each individual channel to 2 nm.

Sensitivity and Accuracy

Integrated in all FV3000 confocal microscopes, TruSpectral detection technology enables much higher light throughput compared to conventional spectral detection units. The volume phase hologram diffracts light with up to three-fold higher transmission efficiency vs. reflection gratings. The result is excellent multicolor fluorescence microscopy images of live and fixed tissue.

Enhanced Quantum Efficiency

The FV3000 microscope’s high-sensitivity detector (HSD) enables you to view samples whose emission is too weak to view with conventional detectors. The HSD unit incorporates two GaAsP channels with a maximum quantum efficiency of 45% and Peltier cooling that reduces background noise by 20% for high signal-to-noise (S/N) ratio images under very low excitation light. HSD units can be combined to enable four-channel GaAsP imaging with the FV3000 system.

Multichannel TruSpectral Detection with Sixteen-Channel Unmixing

TruSpectral detection works independently on all microscope channels, enabling true simultaneous multichannel lambda scanning on up to four channels. Multichannel lambda mode facilitates both live and post-processing spectral unmixing for excellent spectral separation results. With up to four dynamic ranges, bright and dim signals can be optimally separated by independently adjusting the sensitivity of each detector.