What does Prospective Instruments consider a sufficient warm-up time to achieve the frequency-drift specs given for operation without the synchronization option?

After a maximum of 10 minutes from cold start the laser works fine. By simply leaving the temperature control turned on (like a stand-by function, enabled by the Power Key-Switch) stabilized operation will be reached within <5 min.

What is the mean time to failure (MTTF) for the diode pump source?

The FSX-series operates with multiple pump diodes including Telcordia GR-486 core compliant, (i.e. >200'000 hours MTTF), laser diodes reducing pump failure probability.

Are other components more likely to fail before the pump source? What mean time to failure is expected?

Prospective Instruments cannot provide MTTF numbers for the FSX laser. However, all critical components inside the laser are either Telcordia or MIL compliant.

Are there any statistics for maintenance frequency and duration for downtime?

The FSX platform can be serviced in the field, however the system is designed for maintenance free operation over the entire product lifetime. In case of catastrophic failure of a component (e.g., pump laser), Prospective Instruments stocks all long-lead items in-house, ensuring quick response times.

Can the FSX laser with synchronization option also be operated with disabled synchronization option?

Yes. By simply pressing the synchronization button the user can switch the synchronization on or off (it takes a few milliseconds for the Phase-Locked-Loop (PLL) to lock). There is no reset, cold start or anything else required. There will be no pulse drop out or any other pulsed interruption. The pulse repetition frequency will restore to the default value when synchronization is switched off.

Will the operation of the FSX system with synchronization option installed but disabled be identical to the operation of an FSX system without installed synchronization option?

Yes, the operation will be identical. However, for lowest noise operation in free running conditions, the PZT Input of the laser head (18) should be shortened, in order to avoid any noise pick up.

How does the MPX-series work?

The tight integration of the required light engines (femtosecond laser, 8-channel cw light engine, white light source) together with the corresponding detection modules and beam handling optics allows an entirely new way of implementing and using a multi modal microscope. The microscope becomes a truly turn-key, integrated, compact, easy-to-use, maintenance-free and very easy-to-install imaging platform.

The user does not need to worry about laser alignment, detection efficiency or non-compatible or misaligned hardware over time. The entire microscope consists of two parts, controlled through a flexible, non-detachable umbilical, the scanhead and the controller box. The compact scanhead incorporates laser beam handling and scanning, detectors and control electronics. Everything is pre-aligned, rigid and firmly fixed. The scanhead can be rotated and translated in any direction (truly 3D) and enables all multi-photon-based imaging needs. Cells, molecules, even live whole animal imaging can be accomplished.

The controller on wheels incorporates a built-in PC together with other electronic and photonic devices. Only a power cord, keyboard, mouse and display need to be connected to the controller box to make it completely operational. The device can easily be operated even by non-experts. It allows, through the built-in multimodality, extra features and combination of imaging capabilities for correlative microscopy which is not available until now. It is simple to use, and has the utility of a household appliance. Researchers can share the microscope; it is easily moveable and transportable between different locations. This novel capability is unique to the field, and serving a growing need. A modular and flexible product layout allows the user to upgrade the microscope at a later stage and ensures the researcher can add extra features over time on the same platform.

What multi-photon modalities are integrated in your imaging platform?

Currently, you have the possibility to scan one sample in single-photon, two-photon and brightfield modality.

What additional multi-photon modalities are planned to be integrated into the microscope?

We are planning for a FLIM application, and a coherent RAMAN source, as well as a non-coherent one. We are also working on a THG modality at the moment.

What is the imaging workflow of the product?

The user can freely choose between the built-in modalities (1P, 2P and brightfield) to image their samples with a broad spectrum of corresponding modalities. The options are limitless. They can then easily overlay/correlate images and process the multidimensional/multimodal data with our powerful software.

Who can use your product?
  • University labs in the biomedical and neuroscience sector
  • Hospitals with a strong R&D background 
  • The cosmetics, pharmaceutical, diagnostics and clinical CRO (contract research organization) industry sector 
  • Forensic R&D could also be a potential target audience.
  • MDs in the hospitals, working with patients deploying intra operating theatre imaging replacing or enhancing classical HE staining and immunohistochemistry (#instantpathology).

What are your target imaging applications?
  • Label-free imaging
  • Digital pathology
  • Whole slide imaging (WSI)
  • Deep tissue imaging
  • 3D imaging
  • Imaging of fresh biopsies
  • Immunohistochemistry (IHC), Immunocytochemistry (ICC) and other labelling techniques
  • Life cell imaging
  • Organoid imaging
  • Neuroimaging
  • Live animal imaging
  • Whole animal imaging
  • Calcium monitoring

Is the wavelength of the laser mainly designed for SHG/THG? Most customers who are using GFP and GCamp expect the wavelength to be 900-950nm. Is the wavelength of built-in laser selectable?

The laser wavelength is fixed at around 1030nm for now. SHG and 3PEF works, for THG the laser wavelength is still a little bit too short, as THG will be generated too far in the UV range (although it will work in forward direction).

Are there emitting filters for multi-color in 2P-/ two-photon-mode?

Yes, one can use up to 4 PMT channels simultaneously. We typically use Semrock filters (long-pass dichroic and notch filters).

The power of the conventional fs laser is usually >1000mW, is our 100mW power enough?

100mW is indeed on the low side, however with 1000mW in galvo-galvo configuration you will burn the samples.

Does the galvo Scan Type support other resolution modes, such as 256x256?

Yes, we are using ScanImage as scan software. The lowest resolution mode is 64x64.

Is there a resonant scanners option?

Yes, there is the possibility to add a resonant scanner to the basic version.

Does 2P-mode support lens with higher NA?

We have tested up to 0.8NA (Nikon 16x). The higher the NA, the better!

As for 16x and 4x lenses, are they both available in 2P and fluorescence, or only in separate mode?

Our software enables the chose, independent of the modality, of any of the 4 positions on the turret. The Customer can mount any objective on any position and choose it independently.

Does the motorized manipulator support tilting the microscope body and moving along lens for imaging of non-horizontal surfaces and volumes such as temporal lobe observation?

As the scanhead is freely movable (translation & rotation) in all directions, one can move along any axis. However, we don´t have a script that does that automatically. We can deliver such software on demand.

During shipping, will the light path change and require another calibration?

There will be no re-alignment necessary. Our microscope will arrive turn-key out of the box.

Can we overlap images

Yes. Typically, we use ImageJ for that.

What is the highest working space for animal?

There is basically no limit to the working space. The umbilical connecting scanhead and controller is 2 meters long.

How long does it take to image a 1x1cm sample?

We are improving the scanning speed every day. Currently it takes approximately 1 hour to achieve this area.

Does the surface of the sample need to be flat for image acquisition?

Ideally it should be flat. If we get 3-dimensional tissues samples, we gently press them against the glass bottom, of the Fluorodish (or any other) plate.

Do you offer service? How can we get a service technician?

Yes, we do. There is the possibility to sign a service contract. This contract contains: phone support, repair of broken devices and replacement devices. Please contact us for further information.

What is the field of view for 2P-microscopy?

Depends on the galvo-scanangle: You can choose between 60 µm and up to 1200 µm

Can I use conventional fluorescence dyes for 2P-microscopy?

Yes, you can use standard fluorescence dyes and use filter-sets optimized for your spectral range. In addition, you can optimize your experiments by using special commercially available 2P high-absorbance fluorescence dyes.

What is the file-format of the images?

You can save the images as tiff-files and pre-processing the RGB-images (Brightfield) or 16-bit grayscale (1PM and 2PM) images with the image editing software of your choice.

Do I need programming-skills to use your microscopes?

No, we are working with well-established software like e.g. Windows10, ScanImage, µManager, Fiji or MATLAB. All software will be pre-installed and ready-to-use.

How is the software structured?

MATLAB-based, ImageJ and home-built GUI

What is digital staining?

Digital staining means taking advantage of endogenous autofluorescence of certain molecules and structures of your sample, to image and distinguish different tissue and cell architectures and structures, metabolic conditions, pathogenic tissue and so on without the need of external staining’s.

Does the resonant galvo-galvo option also allow scanning at the slower rate offered by the galvo-galvo scanner?

Yes, this is no problem. The customer can choose between galvo-galvo ONLY or with resonant.

Is there a time limitation for continuous recording?

No, there is no limitation. The system is properly air cooled and the power consumption is actually not too high. In fact, our demo device is often under test for a long time without a problem.

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