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FV4000

Applied Technologies
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See Further with NIR-Enabled Confocal Microscopy

The system’s enhanced technologies provided expanded multiplexing to see more in one image.

NIR imaging offers greater multiplexing capabilities by extending the excitation (λ_Ex) and detection (λ_Em) spectral profile of the FV4000 system. This enables additional dyes to be used to help minimize emission signal overlapping.

  • Multiplex up to six channels with updated TruSpectral™ technology and our high sensitivity SilVIR™ detectors
  • High-efficiency volume phase hologram (VPH) grating and slit can detect an industry-leading 400 nm to 900 nm wavelength range* with a minimum 1 nm step
  • Expand your fluorochrome choices with up to six channels of broadband or red-shifted detectors to minimize damage and reduce autofluorescence
  • Modular laser combiners allow up to 10 laser lines from 405 nm to 785 nm in parallel
     

    *As of March 2023.

  • Cell nuclei (DAPI; blue)

    cell membrane (AF488; green)

    nuclear pore (AF561; yellow)

    microtubule (Qdot605; magenta)

    mitochondria (MitoTracker DeepRed; cyan)

    actin (AF750 phalloidin; gray)

    HeLa cells labeled by 6 fluorochromes.

    HeLa cells labeled by 6 fluorochromes.
    Cell nuclei (DAPI; blue), cell membrane (AF488; green), nuclear pore (AF561; yellow),
    microtubule (Qdot605; magenta), mitochondria (MitoTracker DeepRed; cyan), actin (AF750 phalloidin; gray).

High-Quality Optics for Efficient NIR Fluorescence Imaging

The FV4000 system's optical elements have a high transmission from 400 nm to 1300 nm, including the galvanometer and resonant scanner, which are coated in silver rather than aluminum.

Our award-winning X Line™ objectives are corrected for chromatic aberrations between 400–1000 nm. They also have a higher numerical aperture, excellent flatness, and very high transmittance from UV to NIR, increasing the multiplexing capabilities.

For improved colocalization reliability, our specialized A Line™ (PLAPON60XOSC2) oil immersion objective (ne~1.40) significantly minimizes chromatic aberration for strict colocalization analysis.

X Series

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A total of 77 four-channel XYZ positions (11 × 7) were acquired using a 1K resonant scanner within 16 minutes to create the stitched image, which used to require 2 hours using a galvanometer scanner. The coronal section of an H-line mouse brain, cyan; DAPI (cell nuclei), green; YFP (neuron), yellow; Cy3 astrocytes, magenta; AlexaFluor 750 (microtubule). Sample courtesy of: Takako Kogure and Atsushi Miyawaki, Cell Function Dynamics, RIKEN CBS.

High-Quality Confocal Images at High Speed

A unique combination of advanced technologies delivers high-quality images faster than conventional laser scanning microscope systems.

  • High-resolution images at high speed: 1K × 1K resonant scanner at FN20 with 0.033 µs per pixel and the SilVIR detector enable you to rapidly acquire images with minimal noise
  • Exceptional quality macro images: quickly acquire stitched macro images with outstanding quality to maximize your time and research potential

Simple, Precise Super Resolution Imaging

Capture super resolution images using the FV4000 microscope with no dedicated hardware.

  • Easily observe subcellular structures using our A Line HR objectives and our super resolution software (FV-OSR)
  • FV-OSR automatically optimizes the confocal aperture to detect high-frequency components and enhance their contrast to achieve 120 nm resolution
  • Achieve super resolution images 8x faster than previous-generation systems thanks to the SilVIR detector and on-the-fly processing

Confocal mode 1AU (left) versus super resolution mode (right)

High-Resolution 3D Images in Thick Samples

HeLa cell spheroid labeled by DAPI (cyan, cell nuclei) and AlexaFluor790 (magenta, Ki-67). Imaging of the spheroid’s whole volume was possible by NIR 785 nm, although only surface area cell nuclei observation was possible using a 405 nm laser.

When imaging thicker samples, the FV4000 microscope enables you to capture high-resolution, 3D images.

  • Take advantage of NIR’s longer wavelength to penetrate deeper into tissue thanks to the SilVIR detector’s wide dynamic range and sensitivity
  • Image deeper with less scattering and absorption by taking advantage of the fact that light-scattering compounds—like melanin and heme—absorb less light between 700–1500 nm
  • Image significantly deeper than what’s possible with visible lasers thanks to 685 nm, 730 nm, and 785 diode lasers on the FV4000
  • High NA silicone objectives minimize spherical aberration and silicone oil doesn’t dry out, both advantageous for time-lapse imaging
  • Improve overall image quality and Z resolution using TruSight™ deconvolution for stunning 3D images of thick samples
  • Enjoy a seamless workflow from acquisition to publication with specialized cellSens™ software algorithms

Precise Dynamics of Live Cells with Less Damage

Typically, using longer wavelengths for fluorescence excitation for shorter periods of time is better for overall sample health. Using less phototoxic light means you can image for longer periods, enabling you to obtain more consistent and reproducible data from live cell imaging experiments.

The FV4000 system not only provides gentle time-lapse imaging via the 685 nm, 730 nm, and 785 nm lasers, but it also features a dedicated TruFocus Red Z-drift compensator to maintain the focus position. This upgraded TruFocus Red unit supports a larger range of wavelengths and is compatible with a wide range of objectives, including our high-performing X Line™ and A Line™ series.

Time-lapse photo stimulation: the laser injury was performed on C2C12 cells. The green pseudocolor represents the application of an FM 1-43 bath. The image was acquired with a 2 μs galvo scanner and a UPLSAPO60XOHR objective. A 405 nm laser was used for photodamage and a 488 was used to image. Sample courtesy of: Daniel Bittel and Jyoti Jaiswal, Center for Genetic Medicine Research, Children’s National Research Institute.

Time-lapse image of HeLa cells stained with Hoechst33342 (nuclear, blue), MitoTracker Green (mitochondria, green), LysoTracker Red (Lysosome, yellow), SiR-Tubulin (tubulin, magenta), POR-SA-Halo (ER, cyan). Hoechst33342: Ex 405 nm/Em, MitoTracker Green: LysoTrakcer Red: SiR-Tubulin: POR-SA-Halo: Sample courtesy of: Masayasu Taki, Ph.D., Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Japan, Yuichi Asada and Ryusei Aruga, Graduate School of Science, Nagoya University, Japan.

A 17-hour, time-lapse image of HeLa cells stained with MitoTracker Red (mitochondria, magenta), POR-SA-Halo (ER, cyan). MitoTracker Red: Ex 561nm/Em, POR-SA-Halo: Ex 730nm/Em, Sample Courtesy of: Masayasu Taki, Ph.D., Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Japan, Yuichi Asada and Ryusei Aruga, Graduate School of Science, Nagoya University, Japan.

Clear Images at Depth

Use our silicone immersion objectives with the FV4000 microscope and achieve clear images of features and structures deep within your sample. Silicone oil has a refractive index close to that of live cells or tissue, greatly reducing the spherical aberration as compared to air, water, or other oils. With less aberration, you can achieve clearer images of your sample at depth. And silicone immersion oil does not dry out at 37 ℃ (98.6 °F), making it effective for long-term time-lapse imaging.

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