Journal: Aging (Albany NY)

Article Title: Alterations in oxidative, inflammatory and apoptotic events in short-lived and long-lived mice testes

doi:

Figure Lengend Snippet: Panel ( A ) COX2 (72 kDa) expression was evaluated by immunoblotting in testicular homogenates of short-lived (GH-Tg) and long-lived (GHRH-KO and Ames dwarf) mice. Bar plot graphs represent the mean ± S.E.M. and depict the quantification by densitometry of the bands. Results were normalized to actin (42 kDa) and expressed as fold change relative to the control (normal littermates), which was assigned a value of 1. Bar plot graphs represent the mean + SEM; n = 5-7. * p < 0.05, t-Student test. Panel ( B ) PGD2 testicular levels were determined by immunoassay in testicular homogenates from short-lived (GH-Tg) and long-lived (GHRH-KO and Ames dwarf) mice. Bar plot graphs represent the mean + SEM; n = 5-7. * p < 0.05, t-Student test.

Article Snippet: Testicular sections were washed and incubated for 2h at room temperature with biotinylated secondary antiserum (goat anti-rabbit IgG serum, 1:200 for immunodetection of Iba1 and 1:500 for immunodetection of COX2 and StAR from Vector Laboratories Inc., Burlingame, CA, USA) diluted in incubation buffer (2% goat normal serum in PBS for immunodetection of COX2 and StAR or 5% BSA 0.1% Triton prepared in PBS for immunodetection of Iba1).

Techniques: Expressing, Western Blot

Journal: Aging (Albany NY)

Article Title: Alterations in oxidative, inflammatory and apoptotic events in short-lived and long-lived mice testes

doi:

Figure Lengend Snippet: Panel ( A ) COX2-immunoreactive interstitial cells were isolated by Laser Capture Microdissection (LCM) from testicular sections of short-lived (GH-Tg) and long-lived (Ames dwarf and GHRH-KO) mice. The same section is illustrated before (left panel) and after (right panel) LCM. Bar, 25 μm. A total of 50 to 80 COX2-immunopositive interstitial cells were isolated by LCM and subsequently used to evaluate the expression of CD68 (macrophage cell marker) and StAR (Leydig cell marker) by RT-PCR. Panels ( B and C ) Immuno-colocalization of COX2 and Iba1 (macrophage cell marker; Panel ( B ) and immuno-colocalization of COX2 and StAR (Leydig cell marker; Panel ( C ) in testicular sections from a short-lived mouse (GH-Tg) was examined using a light microscope. Bar, 20 (m. Similar images were seen when testicular sections from long-lived (Ames dwarf and GHRH-KO) mice were used (data not shown).

Article Snippet: Testicular sections were washed and incubated for 2h at room temperature with biotinylated secondary antiserum (goat anti-rabbit IgG serum, 1:200 for immunodetection of Iba1 and 1:500 for immunodetection of COX2 and StAR from Vector Laboratories Inc., Burlingame, CA, USA) diluted in incubation buffer (2% goat normal serum in PBS for immunodetection of COX2 and StAR or 5% BSA 0.1% Triton prepared in PBS for immunodetection of Iba1).

Techniques: Isolation, Laser Capture Microdissection, Expressing, Marker, Reverse Transcription Polymerase Chain Reaction, Light Microscopy

Journal: Aging (Albany NY)

Article Title: Alterations in oxidative, inflammatory and apoptotic events in short-lived and long-lived mice testes

doi:

Figure Lengend Snippet: Panel ( A ) the relative expression levels of CD68 and CD163 (macrophage cell markers) and the relative expression levels of StAR (Leydig cell marker) were determined by real time-PCR in testicular homogenates of short-lived (GH-Tg) and long-lived (GHRH-KO and Ames dwarf) mice and their normal littermates. Results were normalized to GAPDH housekeeping gene and expressed as fold change relative to the control (normal littermates), which was assigned a value of 1. Bar plot graphs represent the mean + SEM; n = 5-7. * p < 0.05, t-Student test. Panels ( B and C ) Quantification of Iba1-positive macrophages in testes of GH-Tg short-lived mice, GHRH-KO and Ames dwarf long-lived mice and their normal littermates was evaluated using a light microscope with a magnification of 400x and a gridded eyepiece. Results are expressed as macrophages/mm 2 (Panel B ) and macrophages/tubule (Panel C ). Bar plot graphs represent the mean + SEM; n = 5-7. * p < 0.05, t-Student test.

Article Snippet: Testicular sections were washed and incubated for 2h at room temperature with biotinylated secondary antiserum (goat anti-rabbit IgG serum, 1:200 for immunodetection of Iba1 and 1:500 for immunodetection of COX2 and StAR from Vector Laboratories Inc., Burlingame, CA, USA) diluted in incubation buffer (2% goat normal serum in PBS for immunodetection of COX2 and StAR or 5% BSA 0.1% Triton prepared in PBS for immunodetection of Iba1).

Techniques: Expressing, Marker, Real-time Polymerase Chain Reaction, Light Microscopy

Journal: Nature Machine Intelligence

Article Title: Development of AI-assisted microscopy frameworks through realistic simulation with pySTED

doi: 10.1038/s42256-024-00903-w

Figure Lengend Snippet: a , pySTED is used to confirm the robustness of a model to the random initialization by repeatedly optimizing (50 repetitions) the imaging parameters on the same sequence of data maps (200 images). Two fluorophores are considered for demonstration purposes (Supplementary Table ). b , Resulting imaging optimization objectives from LinTSDiag at three different time steps (10, cyan; 100, grey; 190, red) for 50 independent models, which are presented for increasing signal ratio (top to bottom). With time, LinTSDiag acquires images that have a higher preference score for both fluorophores (purple contour lines) and converges into a similar imaging optimization objective space (red points). c , Standard deviation (s.d.) of the imaging optimization objectives and of the preference scores decreases during optimization (cyan to red), supporting the convergence of LinTSDiag in a specific region of the imaging optimization objective space for both fluorophores. The dashed line separates the imaging optimization objectives (R, resolution; P, photobleaching; S, signal ratio) from the preference network (PN). d , Typical pySTED simulations on two different fluorophores (top/bottom) using the optimized parameters on fluorophore A (left) or B (right). Parameters that were optimized for fluorophore A (top left) result in higher photobleaching and maintain a similar resolution and signal ratio on fluorophore B (bottom left) compared with parameters that were optimized for fluorophore B (bottom right). Supplementary Table lists the imaging parameters. e , Example acquisition of LinTSDiag of tubulin in kidney epithelial cells (Vero cells) stained with STAR RED in the beginning (left) and end (right) of optimization. f , Over time, LinTSDiag manages to increase both resolution and signal ratio of the acquired images (35 images, cyan to red). g , LinTSDiag allows multicolour imaging due to its high-dimensional parameter space capability. LinTSDiag optimizes the averaged resolution and signal ratio from both channels in dual-colour images acquired of the golgi (STAR ORANGE) and NPC (STAR RED) in Vero cells. h , LinTSDiag can maximize the signal ratio in the images and maintain the resolution of the images (35 images, cyan to red).

Article Snippet: Detection of primary antibodies was achieved using secondary STAR RED goat anti-mouse IgG (1:200, abberior, code STRED-1001-500UG) and STAR ORANGE goat anti-rabbit IgG (1:200, abberior, code STORANGE-1002-500UG) for the double staining of NPC and golgi.

Techniques: Imaging, Sequencing, Standard Deviation, Staining