Femto3D-RC of Two-Photon

Real-time 3D Two-Photon microscopy 

Real-time 3D microscopy at Femtonics does not just mean the ability to measure 3D image stacks, but also to collect signals from different depths of the sample fast enough to resolve its functionality, following events all through the three-dimensional space of the sample with a high signal-to-noise ratio. We provide a fast high resolution 3D two-photon laser scanning method (Roller Coaster Scanning) capable of imaging long dendritic segments resolving individual spines and inputs with a temporal resolution of a few milliseconds.






Scanning modes of the microscope


  • RollerCoaster scanning (3D trajectory scanning)
  • Tilted frame scanning (3D arbitrary frame scanning)
  • Stripe scanning (fast line - Z scanning)
  • 4D scanning
  • All 2D scanning modes possible with galvanometer based scanners:
    • Frame scanning,
    • Folded/Multi-frame scanning,
    • Line scanning,
    • Point scanning,
    • XYZ stack

   



In the figure, the dendrite curving out of the focal plane was followed along the 3D trajectory in six dendritic regions and 12 neighboring spines at 150 Hz. Back-propagating action potential induced Ca2+ transients measured in these 18 regions had a signal-to-noise ratio comparable to that of the transients measured in one region by a single line scan using the same conditions as a classical two-photon microscope.

Reference:
G. Katona, A. Kaszás, G. F. Turi, N. Hájos, G. Tamás, E.S. Vizi, B. Rózsa 

Roller Coaster Scanning reveals spontaneous triggering of dendritic spikes in CA1 interneurons, Proc. Natl. Acad. Sci. USA,Volume 108, No. 5 (2011), Page 2148-2153


New 3D measurement modes:

  • Roller Coaster Scanning (3D trajectory line-scanning)
    • 25 μm @ 150 Hz (optimal for calcium imaging)
    • 90 μm @ 40 Hz
    • 100 μm or 300 μm @ 10 Hz (selectable)
  • Tilted frame scanning
    • Frame scanning on planes not parallel to the optical axis
  • Stripe scanning Frame scan
    • Fast line - Z scanning
  • 4D scanning
    • High speed volume scanning  


Using the RollerCoaster scanning you gain:

  • A ~25-fold increase in the access rate to imaging 40-50 μm long continuous dendritic segments
  • Imaging very long dendritic segments (up to 250-300 μm)
  • Resolution of individual spines
  • Simultaneous measurement of 15-50 locations at different depths


Femto3D-RC retains all 2D scanning parameters.


  • Wide field-of-view (e.g. 650 μm × 650 μm with a 20× objective lens)
  • Good spatial resolution characteristic to two photon microscopy (down to 350 nm lateral resolution)
  • Optimized multi-channel non-descanned detection with high efficiency


Femto3D-RC comes with all advantages of the  Femto2D-Galvo microscopes


  • Optimized photon collection due to novel patented technology 
  • (400% higher efficiency compared to our competitors)
  • Real-time data analysis and visualization of 3D measurements
  • Flexible protocol editor
  • Simultaneous optical and electrophysiology recordings
  • Customized microscopes
  • Lab master function to drive lab instruments such as drug delivery system synchronously with 3D measurements.



ROLLERCOASTER SCANNING



Capturing action potential evoked Ca2+ transients on a 250 um long dendritic segment.


STRIPE SCANNING



gCaMP labelled neurons in the mouse cortex responding to visual stimulation. Note that the cells are in different depths and the visible apical dendrites.


MES software 

MES software with the specific extension module seamlessly integrates all new scanning modes into the measurement control and analysis software package.


Contact us Address :Room A020, Floor 3, LianRi International Building, No.18 ,NanLangJiaYuan,ChaoYang District , Beijing City ,100022,China Telephone:010- 65129207 Fax :010- 65129207 Email :tim_liuyi@aliyun.com , tim@hktimwinter.com 京ICP备15043433