Journal: NPJ Precision Oncology

Article Title: Optimized culturing yields high success rates and preserves molecular heterogeneity, enabling personalized screening for high-grade gliomas

doi: 10.1038/s41698-025-00946-1

Figure Lengend Snippet: a Brightfield images of four different representative glioma cultures (Zeiss Axio Observer D1 Inverted Phase Contrast Fluorescence Microscope, 10x magnification). All cultures show an astrocytoma-like phenotype with elongated cell shapes, slim protrusions, small cell bodies, and bright edges. Typically, unless a culture is severely overgrown, there is space between individual cell bodies. The size bar represents 100 μm. b Brightfield images (Zeiss Axio Observer D1 Inverted Phase Contrast Fluorescence Microscope, 10x magnification) of culture GS.0821 at passage 0 and at passage 6. At passage zero, a heterogeneous mix of two cell types can be observed: the astrocytoma-like phenotype and the fibroblast-like phenotype. c Immunofluorescent staining of neuronal stem cell marker SOX2 and GFAP (Leica TCS SP5 microscope). The black and white images show the positive cells for a single marker, the color image shows a composite image. d Percentage of marker-positive cells per arm of the protocol for cell culture GS.0875. Error bars represent differences between three different areas of the same cover slip. e Growth rates of the cultures from the four different arms of the protocol over three consecutive passages for three different tumors. Error bars represent the standard deviation between technical replicates.

Article Snippet: Bright field microscopy images were made on a routine basis with a Zeiss Axio Observer D1 Inverted Phase Contrast Fluorescence Microscope, HAL1 illuminator, and using a 10x Zeiss A-plan objective (421041-9910-000).

Techniques: Fluorescence, Microscopy, Staining, Marker, Cell Culture, Standard Deviation

Journal: Scientific reports

Article Title: Optogenetics design of mechanistically-based stimulation patterns for cardiac defibrillation.

doi: 10.1038/srep35628

Figure Lengend Snippet: Figure 1. Simultaneous all-optical map and control of cardiac conduction pathway. (a) Scheme of the wide-field fluorescence macroscope consisting of a LED at a wavelength centered at 625 nm for excitation of the voltage-sensitive dye and a CW laser at 473 nm for ChR2 activation. The system is provided with a random access scanning head comprised of two orthogonally mounted acousto-optical deflectors (AOD x-y). A 1.25x objective is used for imaging and stimulation of the whole mouse heart, a 20x objective is used to focus the fluorescence signal into a central portion (100 × 100 pixel) of a sCMOS sensor operating at a frame rate of 2 kHz. (b) Electrocardiographic recording of a ChR2-expressing heart, showing sinus rhythm with a frequency of 3 Hz. Blue arrowheads represent the ChR2 stimulation performed at 5 Hz (laser power at the sample 4.9 mW, illumination time 5 ms). The orange line represents the LED illumination. (c) Fluorescence image (F0) of a ChR2-expressing heart stained with voltage sensitive dye (left). Scale bar of 2 mm in yellow. Six representative frames of optical mapping (ΔF/F0) recorded from the same heart. The electrical activation is reported in red and the baseline in cyan. On the right, corresponding color-scaled isochronal map of the action potential reporting the voltage activation time per pixel. (d) On the left, fluorescence image (F0) of a ChR2-expressing heart stimulated in the apex either with an electrode (red spot, above) or with ChR2 activation with the blue laser (blue spot, below). The green frame represents the AODs working-field on the sample. Corresponding optical mapping frames (ΔF/F0) recorded from the same heart are reported next. On the right, corresponding color-scaled isochronal map of the action potential reporting the activation time per pixel.

Article Snippet: The whole mouse heart was excited in wide-field configuration using a 1.25x objective (EC Plan-Neofluar 1.25x/0.03 M27, Carl Zeiss Microscopy) and a LED operating at a wavelength centered at 625 nm (M625L3, Thorlabs) followed by a band-pass filter at 640/40 nm (FF01640/40-25, Semrock).

Techniques: Control, Fluorescence, Activation Assay, Imaging, Expressing, Staining