Light-Sheet Microscopy Tutorial Videos

Theory & Principles - Phototoxicity in Light-Sheet Microscopy

This video shows how Phototoxicity can occur as a result of light exposure during excitation illumination in living samples.

Light-sheet microscopy enables high imaging speed and the possibility of capturing a higher number of subsequent events. This is of particular relevance in the fast-occurring dynamic processes, such as calcium imaging, or when imaging time-lapses of development over long periods of time.

Theory & Principles - Photomanipulation and F-Techniques in Light-Sheet Microscopy

In microscopy, photomanipulation refers to using light to control and modify biological processes within cells or tissues. By employing precise wavelengths, intensities, and exposure durations, biological events can be precisely controlled, providing deep insights into cellular functions and dynamics. All techniques (FRAP, FLIP, FLIM, FRET, FCS, and FCCS) require specific wavelengths. 

Potential photomanipulation applications include:

• Photodamage
• Photoconversion
• Uncaging
• Optogenetics

MuVi SPIM- Octagon Exchange from Cleared Sample to Live Sample Imaging

The video shows the fast and easy transition from Cleared Sample (CS) to Live Sample (LS) imaging with the light-sheet microscope MuVi SPIM from Luxendo.

The transition is easily achieved by exchanging the Octagon, which is the core unit carrying illumination and detection objective lenses, and the sample chamber.

MuVi SPIM - Octagon Exchange from Live Sample to Cleared Sample Imaging

The video shows the fast and easy transition from Live sample (LS) to Cleared Sample imaging with the light-sheet microscope MuVi SPIM from Luxendo.

The transition is easily achieved by exchanging the Octagon, which is the core unit carrying illumination and detection objective lenses and the sample chamber.

MuVi SPIM LS - Sample Mounting

The video shows how to mount a sample (e.g. Zebrafish embryo) in the MuVi SPIM LS from Luxendo.

The steps include preparing the capillaries that will hold your sample, fishing the sample, and inserting the capillary in the sample holder.

The MuVi SPIM features a horizontal setup and is designed for fast imaging of large samples, both living and cleared tissue. The unique 4-axis concept with its two-sided illumination facilitates four orthogonal views of the sample without the need for rotation.

MuVi SPIM CS - Cleared Sample Octagon - Exchange 10x to 20x Objective Lens

The video shows in a few simple steps how to exchange the 20x to the 10x detection lens in the cleared sample octagon.

The Luxendo’s MuVi SPIM CS is an extension of the MuVi SPIM capabilities for 3D imaging of optically cleared samples. The system features an exchangeable cleared sample octagon, which has been cleverly designed to enable flexible configurations of detection lenses (10x and 20x).

MuVi SPIM CS Maintenance - Cleaning the Octagon

The video shows how to maintain the cleared sample Octagon. It is recommended to clean the sample chamber before using it, especially in a multi-user environment. Cleaning the chamber is mandatory when changing clearing solutions.

The Luxendo’s MuVi SPIM CS is an extension of the MuVi SPIM capabilities for 3D imaging of optically cleared samples.

MuVi SPIM CS - Measuring the Refractive Index

The video shows how to measure the refractive index of the clearing solution used in your experiment. This step is necessary to adjust the correction collar of the detection objectives lenses in the cleared sample (CS) Octagon.

The Luxendo’s MuVi SPIM CS is an extension of the MuVi SPIM capabilities for 3D imaging of optically cleared samples.

TruLive 3D Imager - Imaging Dishes

The TruLive3D Dishes are packaged sterile in a pack of six. Each box comes with two packs. Each pack contains 6 dishes, with a total of 12 chambers. Each chamber holds up to 200 microliters of liquid. A total of three dishes can be placed into the TruLive3D Imager. This allows for 6 chambers to be studied at the same time. 

TruLive 3D Imager - Imaging Chamber

The TruLive3D Imager has a benchtop design. Open the incubation lid. From the top view, you see the chamber is filled with water. The TruLive3D Imager has an inverted view with dual-sided illumination. On the sides are two water-dipping illumination objectives. A 25x 1.1 NA detection objective is positioned underneath. The heated lid ensures environmental stability, preventing condensation and evaporation.

LCS SPIM - Sample Mounting

The LCS SPIM is for large, cleared samples. After tissue clearing, sample mounting is accomplished in cuvettes. Different cuvette and holder sizes allow for a wide variety of samples.

LCS SPIM - SPIM Cuvettes

Embedding samples for the LCS SPIM is simple, thanks to cuvettes. Three cuvette sizes allow ideal imaging of differently sized samples.

1. The small cuvette is ideal for one small sample, such as a mouse brain.
2. The medium cuvette is ideal for one medium-sized or multiple small-sized samples.
3. The large cuvette is ideal for one large sample, such as a whole mouse.

To prevent exposure to toxic fumes, the cuvettes can be sealed. Each cuvette size comes with its own lid size.