Journal: bioRxiv

Article Title: Nutlin-3a induces KRAS mutant lung cancer specific methuosis-like cell death that is dependent on GFPT2

doi: 10.1101/2023.05.25.542296

Figure Lengend Snippet: Nutlin-3a induces huge cytoplasmic vacuoles in KRAS mutant NSCLC cells. Cells were treated with nutlin-3a (30 μM) for 6 (d,h) or 24 h (a,b,e-g). A, B Representative live cell images captured by light (a) and holotomography microscope (b) show huge cytoplasmic vacuoles in KRAS mutant but not wild type cells. C Time-lapse video capture images (24 h) of H460 obtained by holotomography microscopy show increase in macropinosomes volume with time and membrane rupturing in the end. D Nutlin-3a induced formation of huge vacuoles were incorporated with dextran (green), but not merged with LysoTracker (red). Representative live images were obtained by confocal laser scanning microscopy (left). Number of macropinosomes were quantitated from the captured image (right) (n≥8). E-G Representative images obtained by confocal laser scanning microscopy. Fixed cells were stained with Rab7 (e), LAMP1 (f), or Rac1 (green), and rhodamine–phalloidin (red) (g) and were counterstained with DAPI (blue). H Western blotting of Rac1 expression. *p<0.05, **p<0.01, ***p<0.001 compared with control. Scale bar: 200 μm (a), 7 μm (b–c), and 20 μm (d–g).

Article Snippet: One normal human bronchial epithelial cell line (BEAS-2B) and ten NSCLC cell lines (HCC827, H1299, H1975, PC9, PC9GR, H820, A427, A549, H460, and H358) were purchased from the American Type Culture Collection (Manassas, VA, USA) and the Korean Cell Line Bank (Seoul, South Korea).

Techniques: Mutagenesis, Microscopy, Membrane, Confocal Laser Scanning Microscopy, Staining, Western Blot, Expressing, Control

Journal: Nature protocols

Article Title: Isotropic Imaging Across Spatial Scales with Axially Swept Light-Sheet Microscopy

doi: 10.1038/s41596-022-00706-6

Figure Lengend Snippet: The illumination source consists of 4 lasers that are combined with dichroic mirrors and focused through a pinhole to improve the beam quality. The filtered laser beam is then enlarged by a beam expander and raised to a 3.5” height above the table with a periscope. A cylindrical lens focuses the laser beam as a line onto the resonant galvo. A folding mirror brings the laser beam to a beam height of 3”. The laser beam passes through a polarizing beam splitter and a quarter-wave plate and is focused via the remote objective onto the remote mirror actuated by a voice coil actuator. The polarization of the returning beam is rotated by 90 degrees by the second pass through the quarter-wave plate and is reflected by the polarizing beam splitter. The laser beam is relayed to the illumination objective, which forms the light-sheet in sample space. The sample is mounted from the top into the sample chamber and positioned by a 3D stage and rapidly scanned in the third dimension by a piezo or voice coil actuator. Fluorescence light is collected by the detection objective and is imaged onto a scientific camera after passing a fluorescence filter.

Article Snippet: 427 nm single-edge laser dichroic beam splitter , Semrock , LM01-427-25 , 1.

Techniques: Fluorescence

Journal: Nature protocols

Article Title: Isotropic Imaging Across Spatial Scales with Axially Swept Light-Sheet Microscopy

doi: 10.1038/s41596-022-00706-6

Figure Lengend Snippet: Equipment List for ctASLM-v1.

Article Snippet: 427 nm single-edge laser dichroic beam splitter , Semrock , LM01-427-25 , 1.

Techniques: