Orca r2 digital camera

Periventricular brain tissue (100 μm thick, as above) was subjected to antigen retrieval consisting of boiling in 10 mM sodium citrate, pH 6, for 10 min followed by three PBS washes, then treatment in 88% formic acid (ThermoFisher Scientific) for 10 min. Tissue was then washed three times in PBS and endogenous peroxidase activity was quenched by incubation in 3% hydrogen peroxide for 10 min. After three PBS washes, tissue was blocked for 1 h at RT in 10% (v/v) milk in PBS followed by incubation for 24 h at 4°C with anti-Aβ42 clone D9A3A 14974 (1:1000; Cell Signaling Technology) in blocking solution. Whole mounts and sections were then washed three times for 10 min each with 0.1% Triton X-100 in PBS and incubated for 1 h at RT with biotinylated anti-rabbit secondary BA-1000, 1:300 (Vector Laboratories). Biotinylated secondary antibody detection was completed using the Vectastain ABC Kit and visualized using the Vector DAB Substrate Kit (Vector Laboratories) according to the manufacturer’s instructions. The sections were imaged using Stereo Investigator software (MBF Bioscience) with a Zeiss Axioskop 2+ microscope (Carl Zeiss MicroImaging) and an Orca-R2 digital camera (Hamamatsu Photonics).

Hut78 cells were seeded at 10⁶/2 mL; treated with 1-µM 8–MOP + 1.6 J/cm² UVA and incubated for 24 h, 48 h and 72 h. Then, ca. 50,000 cells resuspended in phosphate buffered saline (PBS) were placed on a microscopic slide, air-dried and fixed in 4% PBS-buffered formaldehyde for 10 min. Nuclear and peri-nuclear DNA were stained using 1-µg/mL 4′, 6-diamidino-2-phenylindole, dihydrochloride (DAPI; Sigma-Aldrich) solution in water. Images were acquired using an ORCA R2 digital camera and HCImage software (both from Hamamatsu Photonics K.K., Hamamatsu, Japan). Experiment was performed in triplicate. Intact and micronucleated cells were counted manually in five random areas of each slide, and the percentage of micronucleated cells was calculated versus the total cell number.

Worms were mounted onto 2% agar pads and anesthetized with 0.2% levamisole. Nomarski microscopy images were acquired with an Orca-R2 digital camera (Hamamatsu) and either a Leica DMRXA2 microscope or a Zeiss Axioskop 2 plus microscope, driven by Volocity 6.3 software (Improvision, Perkin-Elmer). Images of pathology were analyzed semiquantitatively [13 (link), 57 (link)] (Figures S1A–S1E). For pharynx, gonad and tumor pathologies, images were randomized, examined, assigned scores of 1-5 by two independent scorers, and mean values calculated and rounded. Here 1 = youthful, healthy appearance; 2 = subtle signs of deterioration; 3 = clearly discernible, mild pathology; 4 = well developed pathology; and 5 = tissue so deteriorated as to be barely recognizable (e.g., gonad completely disintegrated), or reaching a maximal level (e.g., large tumor filling the entire body width). Intestinal atrophy was quantified by measuring the intestinal width at a point posterior to the uterine tumors, subtracting the lumenal width and dividing by the body width. Yolk accumulation was measured by dividing the area of yolk pools with the area of the body visible in the field of view at 630x magnification.

Cells were cytospun (Shandon II, Thermo Fisher Scientific), Wright-Giemsa stained, permounted and coverslipped. Images were acquired on a DS-Fi1 camera (Nikon) attached to an Eclipse 80i microscope (Nikon) with a using 40X/0.75 objective.
To examine the morphology of living cells, 10 μl of embryonic blood was loaded between two cover slips mounted onto a glass slide and covered with a third cover slip. Images were acquired with an ORCA-R2 digital camera (Hamamatsu) and NIS-Elements software on an Eclipse 80i microscope (Nikon) with a 40X/0.75 objective. Photoshop CS5 software (Adobe) was used for image processing.