M165 fc stereomicroscope

Zebrafish strains: zebrafish were maintained according to institutional and national ethical and animal welfare guidelines. All experiments were performed under UK Home Office licenses 40/3708 and 70/8588. The zebrafish lines Tg(-26wt1b:EGFP)^{li164 (link)} and tmem33^{sh44343 (link)} were used.
Zebrafish morpholino injections: Embryos were injected at the 1 cell stage. A morpholino complementary to the ATG region of zebrafish pkd2 was injected at 1 ng (5’-AGGACGAACGCGACTGGAGCTCATC-3’)^{42 (link)}. A concentration of 0.4 ng or 1 ng control morpholino (5’- CCTCTTACCTCAGTTACAATTTATA-3’) was injected in these experiments.
Image acquisition and analysis: Zebrafish embryos were imaged at 52–55hpf when cysts became visible, using a Leica M165FC stereo microscope and images were acquired using Leica LASX software. Images were quantified using FIJI v1.52i. Zebrafish renal phenotype analysis was double blinded.

Eye images of 6-day-old female flies expressing Aβ under the control of the GMR-Gal4 driver at 25 °C were taken using a Leica M165 FC stereo microscope. At least 8 flies per genotype were investigated.

The screen for Pid mutants was performed using a Zeiss M2Bio dissecting microscope. Fluorescence images were taken on a Leica DM6000 B upright microscope with 400× amplification. Silencing onset of active transgenes was analyzed using a Leica M165 FC stereo microscope with a 1× objective (120× amplification) or a 4× objective (480× amplification).

Images of histological sections were captured using a Keyence BZ-X810 microscope and software, then processed in Fiji. Live imaging was performed using a Leica M165FC stereo microscope with Leica K5 sCMOS camera and LASX software v3.7.4.23463. Fluorescence imaging was performed using a Leica SP8 confocal microscope with LASX software v3.5.7.23225. The 3-D projections of mpeg1.1:eGFP+ macrophages and PCNA+ cells were generated from z-stacks obtained from 63x confocal imaging using the LASX-3D software v3.5.7.23225.

The Culex quinquefasciatus (California) strain was kindly provided by Anton Cornell (UC Davis), which was originally collected near the city of Merced, California in the 1950s. The Culex quinquefasciatus (Alabama) strain was kindly provided by Nannan Liu (Auburn University), which was collected from Huntsville, Alabama in 2002^{16 (link)}. The mosquitos were reared at 27 ± 1 °C, 75% humidity, and a 12 h light/dark cycle in the insectary room at the University of California, San Diego. The adults were fed with 10% sugar water. After mating, females were fed with defibrinated chicken blood (Colorado Serum Company, # 31142) using the Hemotek blood-feeding system. Egg rafts were collected 4 days after blood feeding. Larvae were fed with fish food floating pellets (Blue Ridge Fish Hatchery, USA). Mosquitos were examined and scored with a Leica M165 FC Stereo microscope with fluorescence. All the work presented here followed procedures and protocols approved by the Institutional Biosafety Committee from the University of California, San Diego, complying with all relevant ethical regulations for animal testing and research. All maintenance and experiments were performed in a high-security Arthropod Containment Level 2 (ACL2) barrier facility. The wastewater and used containers were disposed of by freezing for 48 h, and subsequently discarded as biohazardous materials.