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Super-resolution-enabling detector for functional, live-cell imaging - EMCCD with SRRF Stream

Challenge Background

High-resolution live-cell imaging is a demanding application for several reasons – usually fragile sample, its labelling and a big hurdle of the final stage of the process – the imaging of it. Each of these steps impacts cells’ viability so that the experimenter may be left with only a small number of viable cells that can be ultimately imaged on a microscope. This final step has its own challenges related to the phototoxicity of light and photodamage it causes to the cells and their fluorescently-labelled organelles or compartments.

Labelling cells with fluorophores allows the researchers to delineate structures within the cell and in many cases to track proteins or their complexes en route to various stopovers. Observing fine interplay of all these mechanisms requires optical resolution well below the typical Abbe-limited range of ~200 nm.

To address this, several super-resolution (SR) techniques help unravel the details of this traffic, its cargo and its pathways. However, most SR tools suffer from the fact that they require kilowatts of power per cm2 to achieve 2-3-fold (down to 100-80 nm) improvement in resolution. These levels of power are highly toxic and will limit cause unspecific, unphysiological cellular responses that may not reflect actual processes happening inside a cell.

iXon Ultra

Technology Solution

Electron Multiplying CCD (EMCCD) camera technology with SRRF-Stream is the unique detector solution for live, long (tens of minutes) time-lapse super-resolution imaging. SRRF Stream offers real-time, low-excitation power using conventional fluorophores to deliver prolonged live cell observations and accurate physiology with significantly reduced phototoxicity and photodamage.

Andor Camera Solution for live-cell super-resolution

Andor strongly recommends the iXon Ultra or iXon Life 888 Back-illuminated EMCCD Cameras for live-cell super-resolution experiments. The iXon Ultra and Life are well-established EMCCD platforms for live-cell imaging and their combination with CUDA-enabled nVidia GPUs and MicroManager creates a unique, cost-effective super-resolution solution that doesn’t require a stand-alone, typically complex, costly and phototoxic SR setup.

Date: November 2018

Author: Dr Marcin Barszczewski

Category: Solution Note

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