In order to visualize the ingestion of E. coli by D. pulex, 5 individuals were incubated in mineral water (Volvic) and fed with 106−107 CFU.mL-1E. coli (strain MG1655), preliminarily labelled with 4’,6-diaminophenyl-1H-indole-6-carboxamidine (DAPI) at a concentration of 1 μM. During first trials, food boluses containing E. coli had already reached the distal part of Daphnia guts after 30 minutes. As a result, further feeding experiments were performed during shorter incubation periods of 15, 5 and 2 minutes. Following incubation, D. pulex was narcotized with carbonated water during 1 minute, killed with formaldehyde and mounted on a slide for observation of gut content under an epifluorescence microscope (Olympus, 10x magnification) equipped with a blue excitation filter cube (Olympus, U-MWU, 330–385 nm excitation band).
U mwu
Manufactured by Olympus
Sourced in Japan
The U-MWU is a microscope unit designed for use in various laboratory applications. It provides a stable and consistent platform for microscopic observation and analysis.
Automatically generated - may contain errors
Lab products found in correlation
U mwib, by Olympus (3 mentions)
U niba, by Olympus (2 mentions)
Dp30bw, by Olympus (2 mentions)
U mwig, by Olympus (2 mentions)
Epifluorescence microscope, by Olympus (1 mentions)
Bx50 fluorescence microscope, by Olympus (1 mentions)
Glass bottom, by Greiner (1 mentions)
Orca r2 camera, by Hamamatsu Photonics (1 mentions)
U mwig2, by Olympus (1 mentions)
Cell culture dish, by Greiner (1 mentions)
Aquacosmos, by Hamamatsu Photonics (1 mentions)
Bx51 fluorescence microscope, by Olympus (1 mentions)
7 protocols using u mwu
1
Visualizing E. coli ingestion by Daphnia
2
Fluorescence Microscopy Imaging Protocol
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FISH slides were examined under a fluorescence microscope (Olympus BX-50, Tokyo, Japan) equipped with a cooled charge-coupled device. Images of suitable metaphase spreads were acquired using an Olympus DP70 workstation and the FISH analysis software. The mirror units used for each fluorescence light (FITC, Cy-3, and DAPI ) were U-NIBA, U-MWU, and U-MWIB (Olympus), respectively.
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3
Microscopic Fluorescence Imaging Protocol
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The slides were examined with an Olympus BX-50 fluorescence microscope. Images of suitable metaphase spreads and interphase nuclei were acquired on an Olympus DP70 microscope workstation equipped with a cooled charge-coupled device and FISH analysis software. The miller units used for each fluorescence light (FITC, Cy-3 and DAPI ) were U-NIBA, U-MWU, and U-MWIB (Olympus), respectively.
4
Murine Photoreceptor Cell Viability Assay
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Murine photoreceptor cells (3 × 103 cells/well) were seeded into 96-well plates and cultured at 37 °C. The medium was exchanged for 1% FBS-DMEM, and after 30 min, bilberry extract, or Dp3G, Cy3G, and Mv3G were individually added. The cells were incubated for 1 h. Then 3 mM of DTT was added, and the cells were further cultured for 24 h. At the end of the incubation period, Hoechst 33342 and PI were added to the culture medium (8.1 and 1.5 μM, respectively) and then incubated for 15 min. The cells were photographed through fluorescence filters for Hoechst 33342 (U-MWU, Olympus, Tokyo, Japan) and PI (U-MWIG, Olympus) with a charge-coupled device camera (DP30BW, Olympus). Finally, the cell mortality rate was calculated from the number of PI-positive cells as a percentage of the number of Hoechst 33342–positive cells.
5
Fluorescence Microscopy Visualization of Cellular Proteins
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For fluorescence microscopy, proteins were visualized by fusion to GFP, 3 X mRFP or mCherry. The nuclear chromatin region was stained with bisbenzimide H33342 fluorochrome trihydrochroride (Hoechst 33342). Living cells were observed under a fluorescence microscope (model BX51; Olympus, Tokyo, Japan), and images were obtained by a BX51 fluorescence microscope (Olympus) equipped with an ORCA-R2 camera (Hamamatsu Photonics, Hamamatsu, Japan). Filter sets U-MWU (Olympus), U-MWIB (Olympus), and U-MWIG2 (Olympus) were used for Hoechst 33342, GFP and mCherry/3 X mRFP, respectively. Image acquisition and processing were carried out by using Aquacosmos (Hamamatsu Photonics) and Image J (National Institutes of Health, Bethesda, USA) software. Time-lapse observation was performed as follows. Cells were incubated on SSA at 28°C for 16 hours. After conjugation, cells were inoculated to SSA medium on a cell-culture dish with a glass bottom (Greiner Bio-One, Frickenhausen, Germany) and observed under a fluorescence microscope (model IX71; Olympus) as described in Nakamura et al. (2008) [31 (link)]. Images were processed with Image J.
6
Quantifying Cell Death via Fluorescence
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After 12 h of blue LED light exposure, Hoechst 33342 (excitation, 360 nm; emission, 490 nm) and PI were added to the culture medium at final concentrations of 8.1 and 1.5 μM, respectively, followed by incubation for 15 min. Micrographs through fluorescence filters for Hoechst 33342 (U-MWU, Olympus Co., Tokyo, Japan) and PI (U-MWIG, Olympus) were acquired using a charge-coupled device camera (DP30BW, Olympus). The number of dead cells was determined.
7
Ommatidial Arrangement in Butterfly Eyes
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The spatial arrangement of ommatidial types, both in the ventral and dorsal eye regions, was investigated in the eyes of living butterflies, using epi-illumination with a modified telemicroscopic optical assembly (Stavenga 2002 ) equipped with a 50% beam splitter, a long working distance air objective (LUCplanFLN20X, NA 0.45, WD 6.4-7.6 mm, Olympus, Tokyo, JP), and a monochrome digital camera (Chame-leon3, CM3-U3-31S4M-CS, Point Grey/FLIR BC, CA). The light source, a 500 W Xenon lamp, was bandpass-filtered between 500 and 710 nm (20 nm bandwidth; Asahi Spectra, Tokyo, JP). Ommatidial fluorescence was recorded by replacing the beam splitter with standard fluorescence cubes (U-MWU, U-MWBV; Olympus). Reflectance and fluorescence measurements from four eyes of three females and five eyes of three males, with 100-450 ommatidial measurements per eye, were used to classify the ommatidia into groups using k-means clustering (Hartigan and Wong 1979) (link). After the in vivo measurements, the eyes were fixed, sectioned, and observed under a light microscope. The histology indicated four distinct ommatidial types (dorsal; ventral types I-III), which mapped well with the ommatidial groups derived by k-means clustering.
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