Sponsored Article: New benchmarks for live imaging in your core lab

The careful balance of speed, gentleness & resolution is a prerequisite for live imaging. This balance has now been redefined to new exciting limits

Since the routine use of fluorescent proteins began in the 1990s the drive to image live samples has revolutionized our understanding of modern biology. Live imaging is now a cornerstone of the light microscopy core.

The trade-offs between resolution, speed and gentleness for live experiments are well known. Indeed, aligning expectations of facility users in this regard is critical to avoid the misconception that many frames per second can be teamed with week-long acquisitions in multiple colours, with sub 100nm resolution and in samples 100µm thick.

For any live experiment, the length of acquisition or the number of frames within each acquisition needs to be adjusted to prevent any negative impact of light interaction (1). This can result in the capture of datasets that miss or have insufficient resolution to visualize the key events of interest, or that run for insufficient time to capture the datapoints necessary for robust statistical analysis.

Ideally, providing a range of approaches in the facility that enable 3D acquisitions for longer and/or at higher resolution and in multiple colors would go a long way to shortening this list of compromises. 

The latest technology developments move quickly towards this aspiration.

Lattice light sheet technology hit the headlines in 2014 (2) and is now available on an inverted platform (ZEISS Lattice Lightsheet 7). The automatic alignments provided by Lattice Lightsheet 7 mean that even the most inexperienced of facility users can now access this cutting-edge acquisition approach to capture 3D data of their samples with near isotropic resolution over hours or days at a time (Fig.1). Homebuilt Lattice light instruments have already generated numerous high impact publications and the launch of Lattice Lightsheet 7 sets this approach as the new standard in 3D long term time-lapse imaging. Simply take your petri dish or multiwell plate from your existing imaging system and move it to the Lattice Lightsheet 7 to see the difference for yourself.

In terms of resolution, the push for live super-resolution acquisition is clear from a doubling in the number of publications in the last 5 years. However, restrictions in technology have prevented live, 3D multicolour acquisitions at resolutions below 100nm. Elyra 7 with SIM2 completely changes this and uniquely provides multicolor acquisition with resolution down to 60nm at live imaging speeds (Fig.2). Using standard dyes and fluorescent proteins, SIM2 is the ideal solution for core users seeking an easy approach with the speed and light dose of widefield imaging but with super-resolution results.

Restrictions in acquisition longevity or resolution that previously limited experiments have been removed with the introduction of Lattice Lightsheet 7 and Elyra 7 with Lattice SIM2. These novel technologies pave the way for new and exciting insights in live samples. Sign up to the ZEISS workshops to find out more. 

(1) doi: 10.1002/bies.201700003
(2) doi: 10.1126/science.1257998 

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