SRRF-Stream Real Time Super-Resolution | Andor
Andor

iXon SRRF-Stream

‘SRRF-Stream’ is a NEW real time super-resolution microscopy functionality that operates exclusively on Andor’s iXon Life and iXon Ultra EMCCD cameras.

NEW
  SRRF-Stream Super-Resolution
  • Real Time – enhanced workflow, avoids post-processing. View in ‘Live Mode’.
  • Low Excitation Intensities (mW-W/cm2) – prolonged live cell observations and accurate physiology.
  • Conventional Fluorophores, e.g. GFP – simple labelling, no photo-switching required.
  • Live Cell Dynamics – full FOV super-res images every 1-2 secs. > 10 fps using ROI.
  • Cost-Effective – convert conventional fluorescence microscopes to super-resolution microscopes.
TECHNICAL ARTICLE

iXon SRRF-Stream Comparison

Mitotic Division
SRRF vs SIM
Mitotic Division 2
BCS-40 Membrane
Clathrin Pits
Yeast
Blood
Tubulin

Mitochondria

Image comparison of a fluorescently labelled BPAE cell, recorded with a widefield fluorescence microscope and a SRRF-Stream enabled iXon Life 888 EMCCD camera. A x63 objective was used, with further 2x magnification and 560nm illumination. 100 raw ‘input’ images were recorded for every resultant super-resolution image, resulting in a super-resolution image rate of 0.5 Hz. For a fair comparison without SRRF-Stream, 100 standard widefield images were recorded and then averaged. While the original image was of a larger field of cells, a zoomed ROI of one cell is shown here in order to more easily display a line intensity profile comparison through a small region. The improvement in resolving power is readily apparent.

iXon SRRF-Stream Applications

With its ability to smash through the classical diffraction limit, and furthermore, to accomplish this in real time, with non-complex sample labelling, conventional equipment and with low intensity illumination, SRRF-Stream paves the way to unlock previously unseen cellular structure and behaviour, at unprecedented spatio-temporal resolution in a low-photodamage friendly manner.

Observational Capabilities of SRRF-Stream

  • Elucidation of protein structure analysis at a sub-organelle level
  • Tracking of single molecules inside cells
  • Using this tracking to gain insight into individual molecular machinery underpinning cellular physiology
  • With this new information, update models of cell function

Applications of SRRF-Stream

  • Membrane fusion involving individual SNARE protein machinery
  • Dynamic changes of and within synaptic vesicles
  • Dendritic spines reformation due to synaptic plasticity and learning
  • Signal transduction processes and cell-to-cell communication and differentiation

iXon SRRF-Stream Workflow Advantage

Adopting the recently developed SRRF technology from the lab of Dr Ricardo Henriques, University College London (UCL), and working in close collaboration with Dr Henriques, Andor have enhanced the technology to run optimally on iXon EMCCD cameras. Andor are also expert in advanced GPU processing optimization techniques, employed in this instance to execute the SRRF algorithm up to 30x faster than the existing ImageJ-based post processing implementation of SRRF (NanoJ-SRRF). This significant acceleration enables workflow enhancement, by allowing data acquisition and SRRF processing to operate in parallel.

Processing Speed Comparison - SRRF Stream vs NanoJ-SRRF

  • Input image pixels - 1024 x 1024
  • # input images per SRRF image - 100
  • Output SRRF pixels - 4096 x 4096
  • NVidia GTX 1080 GPU card

This graph compares the rate of processing of blocks of 100 raw input images (1024 x 1024 pixels), to yield resultant SRRF super-resolution images of 4096 x 4096 pixels. SRRF-Stream is compared to NanoJ-SRRF, the processing occurring on the same Nvidia GTX 1070 GPU card. The SRRF-Stream acceleration subsequently allows data acquisition and processing to happen in parallel, yielding a further workflow improvement over NanoJ-SRRF.

Since processing is now much faster than the camera can acquire data, ‘SRRF-Stream enabled’ cameras now accomplish real time super-resolution, with large field of view super-resolution images.

Having thoroughly tested SRRF-Stream in our own lab, we are very impressed by both the workflow and also the ability to now utilise larger fields of view for live cell super-resolution. By seamlessly combining the SRRF algorithm with the high-performance of the iXon, we have accomplished the world’s first super-resolution camera for fluorescence microscopy.

Dr Ricardo Henriques, Quantitative Imaging and Nanobiophysics Group, UCL

iXon SRRF-Stream Accessibility

What Do I Need?
MicroManager



What do I need to access SRRF-Stream?

  • A NEW iXon Life or iXon Ultra EMCCD camera
  • SRRF-Stream license (unlocks SRRF-Stream in camera)
  • A CUDA-compatible NVidia GPU card*
  • MicroManager software or Andor SDK2

* The Nvidia GPU card should have Compute Capability v3.0 or above and 4GB or greater on-board GPU RAM. Note that Andor have done a lot of testing using the ‘mid-range’ GTX 1070 and found that, with SRRF-Stream, it is more than adequate to process data much faster than the rate of iXon data acquisition.

Additional Resources

The Learning Center hosts a wide range of tutorial videos, technical articles and webinars to guide you through the range of products for all your imaging needs. We have provided some links below which will get you started on some of our most recent uploads.

  • Technical Article: SRRF-Stream
  • Webinar: NanoJ-SRRF & SRRF-Stream: Fast Live-Cell Conventional Fluorophore Super-Resolution for Most Modern Microscopes