Scientifica HoloStim-3D | SLM Module for Multiphoton Photostimulation
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    The HyperScope multiphoton imaging system now has advanced imaging capabilities; the introduction of an extended wavelength lens set means you can image deeper and through thin scattering layers in in vivo samples. Learn more here.

    Scientifica HoloStim-3D

    Interrogate larger neural networks at higher resolution than ever before, with the world's highest performing multiphoton all-optical solution.

    The HoloStim-3D seamlessly integrates with the HyperScope, an award-winning multiphoton imaging system, to create an industry-leading spatial light modulator (SLM) system for all-optical interrogation of neural networks with previously unachievable performance.

    Investigate neuronal circuits with 3D precision

    The HoloStim-3D enables you to perform more complex photostimulation experiments than those that are possible with random-access two-photon or widefield photostimulation.

    Discover the roles of neuronal subtypes and brain circuits by simultaneously targeting multiple cells with cellular resolution.

    You can observe and record comprehensive network activity by combining precise cell stimulation and imaging with the HoloStim-3D.

    By selectively photostimulating only the target cells and rapidly switching between stimulation patterns, you can better understand neuronal interactions in networks.

    Benefits

    Selective and accurate targeting

    The industry-leading resolution of the Holostim-3D allows cell bodies and subcellular structures to be specifically targeted with minimal off-target activation. This gives you cleaner, more accurate data that you can draw conclusions from.

    Analyse larger neuronal networks

    Target cells in larger brain regions to understand larger neuronal networks. The large field-of-view of the HoloStim-3D also allows you to use a higher magnification and resolution objective and still achieve the required field-of-view.

    Freely combine different imaging and stimulation wavelengths

    Being able to excite or image target cells expressing different opsins or fluorescent proteins increases your experimental flexibility.

    Perform simultaneous two-photon imaging and photostimulation

    Activate and image hundreds of individual cells simultaneously to interrogate neuronal circuits and identify the role of neural subtypes in brain circuits and behaviours.

    Deeper imaging and stimulation

    Specifically target cells and subcellular structures in layer 4 and beyond, achieving high-quality images.

    Mimic neuronal firing

    You can quickly switch stimulation patterns to better mimic neuronal firing, improving the accuracy of your data.

    High laser transmission

    Keep your costs to a minimum by activating the same number of cells with a lower power laser.

    User-friendly design

    The modular design means the HoloStim-3D is seamlessly integrated with the Scientifica HyperScope, a proven multiphoton imaging system, so you can easily upgrade to SLM capabilities and start generating high-quality data faster.


    Mouse layer ii/iii neurons expressing GCaMP6s (calcium indicator) and C1V1 (red-shifted channelrhodopsin) are simultaneously stimulated using the HoloStim-3D, while their calcium responses are imaged using resonance two-photon imaging on the HyperScope. In the total 100 cells were stimulated at 6 nJ per cell, using a spiral pattern. Data acquired by Kelly Sakaki (Scientifica), Robert Lees and Adam Packer (both University of Oxford).

    HoloStim-3D on the HyperScope 360° view

    Example data

    Fig. 1. (a) HoloStim-3D targeted cells within a mouse brain, with cells transfected via viral vector, and expressing GCaMP-6S and C1V1 for photostimulation. In the total FOV (745 μm), 100 cells were selected within an area of 450 μm and targeted for photostimulation with a spiral spot of approximately 6 mW/cell. (b) Each cell is identified with the target label and the corresponding activity observing during imaging with the resonant scanning mirror is shown in the line plots in Fig. 1c,d. Simultaneous photostimulation of each cell was activated 10 times following a 10 s lead-in over a total time of 110 seconds. The photostimulation was activated for 250 μs and the power was controlled via software control connected to a total power control (TPC) module onboard the Amplitude Satsuma (20 W output). Data acquired by Kelly Sakaki (Scientifica), Robert Lees and Adam Packer (both University of Oxford).
    Fig. 2. Using most of the same conditions as the results seen in Fig. 1(a-d), the SLM targeted cells within a mouse brain, with cells transfected via viral vector, and expressing GCaMP-6S and C1V1 for photostimulation. In this example, 21 cells were selected within an area of 250 μm and targeted for photostimulation with a spiral spot of approximately 6 mW/cell. Data captured in-vivo using Scientifica's HyperScope RGG-GG multiphoton imaging system with HoloStim-3D and a 16x objective. Data acquired by Kelly Sakaki (Scientifica), Robert Lees and Adam Packer (both University of Oxford).
    Fig. 3: All-optical interrogation of neural circuits: neurons are stimulated by multiple beamlets that draw spirals over the soma. Activated neurons generate action potentials.

    Software

    Adopting SLM technology is made easier with full integration of globally-recognised ScanImage software, saving you time so you can start experimenting faster.

    Design & Specifications

    Wavelength range standard
    Wavelength range standard
    920 - 1064 nm
    Wavelength range optional
    Wavelength range optional
    700 - 1300 nm
    Stimulation point resolution
    Stimulation point resolution
    <2.5 microns
    Array size
    Array size
    17.7 x 10.6 mm
    SLM response time
    SLM response time
    3.3 ms
    SLM array resolution
    SLM array resolution
    1920 x 1152 pixels

    Accessories

    Filter Cube Module

    A compact, flexible and efficient epifluorescence illuminator, optimised for use with LED light engines. Specifically designed for Scientifica microscopes, the FCM ensures optimal illumination of samples in all possible configurations.

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