Salmonella typhimurium is a well-known bacterium which infects human by entering the epithelial cells of the small intestine and the underlying connective tissue. This causes gastroenteritis, a debilitating disease that can lead to abdominal pain, vomiting, diarrhea and, if not sufficiently treated, death.
S. typhimurium is known to manipulate the host cytoskeleton. To have a closer look, MDCK epithelial cells were infected with the bacterium (green). The images acquired by the THUNDER Imager 3D Cell Culture enable researchers to see an extensive actin (red) rearrangement in the infected cells.
Raw images vs. Large Volume Computational Clearing. Courtesy of Dr. Mark Jepson, Bristol University (UK).
Infection with rabies virus, without vaccination protection or post-exposure prophylaxis, is frequently deadly. The Rabies virus can be transmitted via the mucus or saliva of infected animals. Starting from a bite wound, it moves along neurons to the central nerve system.
The THUNDER Imager 3D Cell Culture was used to image a 450 µm z-stack in 2 channels within 4 minutes to identify the viral phosphoprotein (red) inside the infected neurons of a DISCO cleared ferret brain (nuclei in blue). The spread of the rabies virus inside the animal brain was easily visualized with the THUNDER Imager 3D Cell Culture.
Raw images vs. Large Volume Computational Clearing. Courtesy of Dr. Stefan Finke, Friedrich-Loeffler-Institute, Riems (Germany).
The Influenza virus enters animals via the mucous membrane and infects the respiratory tract. A typical pathogenesis can include fever, dry cough and body aches. Complicated super-infections with bacteria can be life threatening.
The THUNDER Imager 3D Cell Culture visualizes an influenza (red) infected primary porcine lung epithelial grown in a multilayer of about 60 µm. The top layer shows cilia (green), which are responsible for mucus transport. Nuclei are stained in blue.
Raw image vs. Large Volume Computational Clearing. Courtesy of Dr. Stefan Finke, Friedrich-Loeffler-Institute, Riems (Germany).
Key Advantages of the THUNDER Imager 3D Live Cell & 3D Cell Culture
Our product managers talk about key advantages that the THUNDER Imager 3D Live Cell & 3D Cell Culture offer.
With the THUNDER Imager Live Cell, explants of abdominal aorta were imaged using a 20x, 0.4NA long working distance objective. The explants were loaded in well-plates with a thin layer of growth media covering them. A stage top incubator was used to ensure a temperature of 37 Celsius and a humidity of 95%. Four samples, separated by a large distance (> 1 inch), were imaged via LAS X Navigator with custom regions for each explant. To ensure the cell motion was captured, each region had a custom z-stack defined locally, ensuring each explant was in-focus. The total capture time was 2 minutes and 20 seconds and a cycle time of 20 minutes was used for a time-lapse of 46-hours (139 cycles). Additionally, adaptive focus control (AFC) was used to lock the objective to the sample, keeping seamless focus for the entire time-lapse even with large XY movement to address each sample. Instant Computational Clearing (ICC) from Leica Microsystems was applied to the imaged regions to clear out-of-focus haze. This greatly improved the ability to see the fluorescence from the explant tissue as well as the cell migration.