Lattice Light-Sheet Microscopy for Department of Chemistry

The instrument will be in the department of Chemistry in the group of Nikos Hatzakis and will be involved in the center for 4D cellular dynamics

and the challenge center for optimized oligo escape. The main scope of this work is to observe in real time and in 4D (3D and time) live cells and how ligands or nanocarriers, or Lipid nanoparticles with oligonucleotides or other nanomaterials are entering and being delivered in cells. A central part of the work is parallel recording of multiple cells, biocompatibility with imaging slides and using as little volume of material as possible.

The intended procurement is Zeiss lattice light-sheet microscopy. It is an automated, and easy-to-use system, allowing live cell imaging at subcellular resolution with best protection from photo damage.

The adaptability of the instrument allows using multicell well plates (up to 96 well plates) offering parallelized imaging at diverse conditions. We will use this microscopy to study cell dynamics in 4D. Meanwhile, it will be open to other research group, allowing researchers to volumetric imaging of subcellular structures and dynamics.

The instrument of Zeiss has the unique capability of inverse optical configuration where the two objectives are placed below

the sample slide (akin to inverted microscopes). Alternative solution from competitors has the two objectives coming from top and therefore need to be submerged into the solution for imaging.

The Zeiss configuration is necessary for the following two reasons:

a) it allows the imaging samples with minimum volume of 100 uL, while other lightsheet microscopes need minimum volume of~ 2 mL or more. The 20 times volume difference make it prohibitively more expansive for use on a series of experiments that

are central for my research and some of the centers research that includes the addition of very expensive ligands on cells.

b) Compatibility with all cell chamber slides. A big part of our studies require cells imaging in multi-well plates. These can contain from 8 – 96 wells that allow parallel (or very fast) imaging between multiple wells, each well representing different experimental condition. Zeiss can

accommodate 8 well plates and up to 96 well plates .

c) The inverse optical configuration allows adding on top of the slide the T12 transwell cell culturing insets Transwell® polycarbonate

membrane cell culture inserts alow us to image epithelial cells on the proper orientation on the microscope. This is central for some of my people and collaborators work.

There is service in house in Denmark from Zeiss.

Several elements would have to be changed if using another supplier not offering the inverse configuration and compatibility of incompatibility or disproportionate technical difficulties in operation and maintenance from Zeiss.

A) A central part of our work involves imaging with small volumes . This is essential for keeping the cost low, as most of the material and ligands are very pricy. We would be unable to perform experiments at the same cost.

B) Reduce compatibility with most chamber slides. A big part of our studies require cells imaging in multi-well plates able to

accommodate 8 – 96 wells. This allow parallel (or very fast) imaging between multiple wells, each of which constitutes a

different experiment. Without the inverse configuration we would acquire data one experimental condition at time, increasing

multiple times the time required for experiments.

C) challenge imaging epithelial cells on the proper orientation on the microscope. Using T12 transwell cell culturing insets allows my

group to modify their physical dimensions and add them on top of the microscope slide directly. This in turn allows us to image

epithelial like cell on proper orientation. While this is a small part of my group any other configuration would not allow us to do

this type of measurements.