Male flies (n = 30 per replicate, 6 to 10 replicates per genotype) were kept on diuretics, different sugar diets, and high-ion diets for 5 days before they were collected and euthanized. The following diuretics and control diets were used: furosemide (25 mg/ml, Sigma Aldrich) dissolved in acetone (Sigma Aldrich), amiloride (25 mg/ml, Sigma Aldrich) dissolved in water, mannitol (25 mg/ml, Sigma Aldrich) dissolved in water, and acetone-supplemented food. The sugar diets were made with either 40 and 10 g sucrose or glucose (Sigma Aldrich). All sugar diets contained 10 g yeast (protein, Merck Millipore) and were made on deionized water. Diets enriched in NaCl (0.1 mg/ml, Sigma Aldrich) and KCl (0.1 mg/ml, Sigma Aldrich) were made by adding the ion solution to the standard Jazz mix food. Flies raised on untreated standard Jazz mix food were used as control. Body and hemolymph weights were measured as described previously, and photos were taken with a Leica M125 microscope with a ProgRes C14 plus camera (Jenoptik) and the ProgRes CapturePro 2.8 Jenoptik Optical system. The images were handled using ImageJ, Fiji edition (Schindelin et al., 2012 (link)). Graphs represent mean (±SD), and differences were calculated with Kruskal–Wallis and Dunn’s correction (∗p < 0.05, ∗∗p > 0.01, ∗∗∗p > 0.001) using GraphPad Prism, version 5.
Progres c14 plus camera
Manufactured by Jenoptik
Sourced in Germany, Japan
The ProgRes C14 plus is a digital microscope camera that captures high-quality images. It features a 1.4-megapixel CMOS sensor and supports resolutions up to 1392 x 1040 pixels. The camera is capable of providing live images with frame rates up to 30 frames per second.
Automatically generated - may contain errors
Lab products found in correlation
M125 microscope, by Jenoptik (2 mentions)
Progres capturepro software, by Jenoptik (2 mentions)
Pierce ip lysis buffer, by Thermo Fisher Scientific (2 mentions)
Amiloride, by Merck Group (1 mentions)
Mannitol, by Merck Group (1 mentions)
Furosemide, by Merck Group (1 mentions)
Acetone, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Capturepro 2, by Jenoptik (1 mentions)
Bx53 microscope, by Jenoptik (1 mentions)
Axioscope 7 microscope, by Zeiss (1 mentions)
Zen software, by Zeiss (1 mentions)
Lsm 780, by Zeiss (1 mentions)
Bx60 microscope, by Olympus (1 mentions)
Tissue tek oct compound, by Electron Microscopy Sciences (1 mentions)
Vectashield mounting media with dapi, by Vector Laboratories (1 mentions)
Bx53 microscope, by Olympus (1 mentions)
Pierce protease and phosphatase inhibitor mini tablet, by Thermo Fisher Scientific (1 mentions)
β galactosidase reporter gene staining kit, by Merck Group (1 mentions)
8 protocols using progres c14 plus camera
1
Fly Diuretic and Ion Regulation
2
Imaging Malpighian Tubules in Flies
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Flies were quickly frozen in −80°C and imaged using a Leica M125 microscope with a ProgRes C14 plus camera (Jenoptik) and the ProgRes CapturePro 2.8 Jenoptik Optical system. The Malpighian tubules from female and male adult flies were dissected and mounted in 50% glycerol before imaging. Images were handled using ImageJ, Fiji edition (Schindelin et al., 2012 (link)).
3
Examining CG18549 Knockdown Effects
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F1 progenies from da-GAL4 > w1118 (Driver ctrl), w1118 > CG18549 RNAi (RNAi ctrl) and da-GAL4 > CG18549 RNAi (CG18549 knockdown) were collected and examined. Both CG18549 RNAi lines were used, and the counting was performed on progenies from three crosses of each genotype. The mean (±95% CI) are plotted for male and female progenies separated as well as merged for each RNA line. Differences were analyzed using Kruskal–Wallis with Dunn’s comparison. Progenies were investigated for developmental flaws using a Leica M125 microscope with a ProgRes C14 plus camera (Jenoptik) and the ProgRes CapturePro 2.8 Jenoptik Optical system.
4
Fungal Species Identification Protocol
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A total of 95 specimens were subjected to the identification based on the macroscopic and microscopic characteristics (Table 1 ). Dried materials mounted in distilled water, 5% KOH, 1% phloxine, Melzer’s reagent and Congo red using a model of Olympus BX53 microscope and Jenoptik ProgRes C14 Plus Camera (Jenoptik Corporation, Jena, Germany). Microscopic parameters were measured using ProgRes Capture Pro software version 2.8.8. (Jenoptik Corporation).
5
DIC Imaging and Lipid Droplet Analysis
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Differential interference contrast (DIC) imaging was done for all replicates from the table with an Olympus BX-60 microscope (Olympus, Japan) with a ProgRes C14plus camera and the ProgRes CapturePro Software (version 2.9.01) (JENOPTIK AG). The morphology of chosen conditions (Fig. 1 , Extended Data Figs. 4 –6 and Supplementary Fig. 1 ) of Mesotaenium cells that were 89 h on the table was analysed.
For algae that were used for quantifying the abundance of LD per cell, a ZEISS Axioscope 7 microscope (Carl Zeiss) was used including the Zen software (Carl Zeiss). The LD count was carried out in Fiji159 (link). For statistical analysis of the LD count data, we first used a Shapiro–Wilk test160 (link) to assess normality and used Mann–Whitney U tests161 (link) with R (version 3.6.1) accordingly.
Confocal laser scanning microscope was done on a Zeiss LSM780 (Carl Zeiss) set as in Müller et al.162 (link). For the staining of the LD structures, we used the neutral lipid specific stain BODIPY 493/503 (EM/EX) (Merck). Mesotaenium cells were grown for 22 days on WHM medium at 70–80 µmol photons m−2 s−1 and 22 °C. These cells were ultrasonicated for 1 min with 1:500 BODIPY and incubated on a shaker for 5 min before visualization.
For algae that were used for quantifying the abundance of LD per cell, a ZEISS Axioscope 7 microscope (Carl Zeiss) was used including the Zen software (Carl Zeiss). The LD count was carried out in Fiji159 (link). For statistical analysis of the LD count data, we first used a Shapiro–Wilk test160 (link) to assess normality and used Mann–Whitney U tests161 (link) with R (version 3.6.1) accordingly.
Confocal laser scanning microscope was done on a Zeiss LSM780 (Carl Zeiss) set as in Müller et al.162 (link). For the staining of the LD structures, we used the neutral lipid specific stain BODIPY 493/503 (EM/EX) (Merck). Mesotaenium cells were grown for 22 days on WHM medium at 70–80 µmol photons m−2 s−1 and 22 °C. These cells were ultrasonicated for 1 min with 1:500 BODIPY and incubated on a shaker for 5 min before visualization.
6
Histological Analysis of Cardiac Tissue
7
Fungal Specimen Collection and Identification
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The 318 specimens used in the present study are listed in Table 1 , and dried specimens were deposited in the herbarium of Korea National Arboretum (KH). The specimens were collected during June, September, and October 2012 and in June and September 2013. Macro-morphological characterstics were based on field notes and color photos of basidiomata. Micromorphological characteristics were obtained from the dried specimens after sectioning and rehydrating following the mothod of Largent et al. [14 ]. Microscopic observations were made using an Olympus BX53 microscope (Olympus, Tokyo, Japan) and a Jenoptik ProgRes C14 Plus Camera (Jenoptik, Jena, Germany). Measurements of microscopic characters were made using ProgRes Capture Pro v.2.8.8. software (Jenoptik). Morphological identifications were made based on reliable publications [15 16 ]. The current scientific names of the collected specimens were checked at the Index Fungorum (http://www.indexfungorum.org/Names/Names.asp ) or MycoBank (http://www.mycobank.org/defaultinfo.aspx?Page=Home ).
8
β-Galactosidase Reporter Assay in Genetically Modified Mice
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One mouse from each genotype (WT, HET and KO) and sex was euthanized through cervical dislocation, and the brains were dissected and divided into coronal sections by using a brain matrix. Sections were stored in Pierce™ IP lysis buffer and Pierce™ protease and phosphatase inhibitor mini tablets (Thermo Fisher Scientific, Uppsala, Sweden) at 4 °C for 2 h and thereafter used for β-galactosidase staining. The β-galactosidase reporter gene staining kit (#GALS, Sigma-Aldrich (St. Louis, MO, USA)) was used to verify the knockout construct (Figure 1 A). Briefly, the sections were washed twice in 1 mL of PBS, fixed in 1 mL of fixation solution for 25 min, washed twice with PBS and incubated at 37 °C for 2 h with 1 mL of staining solution. Image acquisition of the brain sections was performed with a Leica M125 microscope with a ProgRes C14 plus camera (Jenoptik) and the ProgRes CapturePro 2.8 Jenoptik Optical system.
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