LSM Lightfield 4D

Life moves. Many neuronal and physiological processes occur at very high speeds, making it difficult to accurately capture their spatiotemporal dynamics. Although established technologies have become faster, the required acquisition time still increases with sample volume, so fast processes like neuronal activity or heartbeats require a trade-off between volumetric information and image frame rate. With Lightfield 4D, you no longer have to compromise, as you can capture 80 volumes per second without time delay in 3D. This makes it possible to follow neuronal activity in zebrafish brains, track tissue movement in developing Drosophila embryos, and keep track of moving structures in crawling C. elegans larvae. The unique one-snap-one-volume imaging ensures that crucial events are not missed or distorted. Highly time-resolved particle tracking in complete volumes is finally possible. Start your experiments immediately – on your confocal and without the need to adjust sample preparation.

Collecting 3D information of living samples has always been a challenge, especially for large sample volumes. Optical sectioning requires sequential acquisition of single images to create a Z-stack. Each slice requires light exposure, which is not fully limited to the plane of illumination and can easily add up to harmful amounts across the volume. Lightfield 4D works differently: A complete Z-stack is acquired with a single illumination event, reducing light exposure and phototoxic effects to a minimum. Living samples can be imaged over long periods of time at high temporal density. This combination of outstanding 3D imaging speed and extreme gentleness allows you to follow the sample in multicolor over time without influencing the recorded living activity. You can observe developmental processes, cell migration, vesicle movement or other changes in tissues and organisms that take hours or even days to complete, and still achieve the temporal resolution needed to understand the dynamics.

Typically, acquisition time of large volumes is the critical factor that limits the throughput of imaging. Acquiring a large volume with a single image snap speeds up your experiments by multitudes. The unmatched speed with which Lightfield 4D captures multi-color volumes can be used to increase the productivity of experiments in a variety of ways: Image and analyze more samples than ever before in every session, immediately improving experimental statistics. Compare multiple different sample cohorts of wild type and genetically modified phenotypes, or samples with different drug treatments. Instead of hours, only minutes are spent collecting the data you need, leaving you more time for advanced analysis and investigation of your datasets.

Laser scanning microscopes (LSM) have proven to be the most versatile microscopy systems. They combine super-resolution and spectral imaging with high-quality optical sectioning of large samples, along with the capability to incorporate additional fluorescent information and molecular dynamics measurements. Take your experiments to the next level by pairing this remarkable flexibility with the gentle and instant volume imaging of Lightfield 4D:

• Monitor neuronal activity in 3D at high speed, and supplement this with super-resolution structural details captured with Airyscan.
• Track macrophage movement during a wound healing assay and add high-resolution details of the wound site to your investigation.
• Leverage the photomanipulation capabilities of your LSM for bleaching, photoactivation, photoconversion, or ablation experiments, followed by gentle volume imaging.
• Accomplish all of this on the same microscope as part of the same experiment without ever moving your sample.

To truly capture the essence of biological processes, imaging must be done in 4D, as both volume and time are essential for investigating living systems. This concept is not new; many optical sectioning techniques have been developed over the past decades to attempt to meet this requirement. However, these methods typically rely on sequential image acquisition to create Z-stack images of volumes, which introduces time differences within the sample volume, severely limiting the imaging speed and the spatiotemporal accuracy of the acquired data.

Lightfield 4D offers a unique solution by imaging an entire volume at an exact point in time, without any time delay. Instead of capturing single 2D images at different time points, a micro lens array positioned in between objective and camera generates 37 individual images, collecting all of the 3D information at the same instant. Each of these different views provides both spatial and angular information which serves as the foundation for creating a Z-stack through deconvolution-based processing. In this way, Lightfield 4D can generate 80 volume Z-stacks per second.