Three RNAs encoding LUC and EGFP were synthesized and purified as previously described43 (link). Briefly, pCS2 plasmids, pCS2-LUC-EGFP-pA, pCS2-nt5ea-LUC-EGFP-pA, and pCS2-nt5eb-LUC-EGFP-pA (Fig. S1 a,b), were linearized with NotI and purified using the NucleoSpin Gel and PCR Clean-up kit (Macherey-Nagel) for in vitro transcription. The capped RNA was transcribed from the purified DNA templates using the Message mMachine SP6 Kit (Life Technologies, Carlsbad, CA, USA). The synthesized RNA was purified using an RNeasy Mini Kit (Qiagen) for microinjection. Approximately 2–4 nL of 100 ng/µL of each RNA was injected into the cytosol of embryos at the one-cell stage, as previously described44 (link). The injected embryos were observed under an SZX16 fluorescence stereomicroscope with the SZX2-FGFP filter set (Olympus, Tokyo, Japan). Microscopic images were captured using a DP73 camera and cellSens image acquisition software (Olympus).
Cellsens image acquisition software
Manufactured by Olympus
Sourced in Japan
CellSens is Olympus' image acquisition software designed for microscopy applications. The software facilitates the capture, processing, and management of digital images from Olympus microscopes. It provides essential tools for controlling microscope hardware and acquiring high-quality images.
Automatically generated - may contain errors
Lab products found in correlation
Message mmachine sp6kit, by Thermo Fisher Scientific (1 mentions)
Szx16 fluorescence stereomicroscope, by Olympus (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Nucleospin gel and pcr clean up kit, by Macherey-Nagel (1 mentions)
Dp73 camera, by Olympus (1 mentions)
Eclipse e600 microscope, by Nikon (1 mentions)
Accutase, by Thermo Fisher Scientific (1 mentions)
Bx53 microscope, by Olympus (1 mentions)
Hematek 3000 system, by Siemens (1 mentions)
Dp72 digital camera, by Olympus (1 mentions)
Bx52 microscope, by Olympus (1 mentions)
Masson s trichrome staining, by Abcam (1 mentions)
Cm3050, by Leica (1 mentions)
Oct medium, by Thermo Fisher Scientific (1 mentions)
Superfrost plus glass slide, by Thermo Fisher Scientific (1 mentions)
Bx51 microscope, by Olympus (1 mentions)
Sz61 stereomicroscope, by Olympus (1 mentions)
Sc30 camera, by Olympus (1 mentions)
Model eclipse ni u, by Nikon (1 mentions)
Dp 80 dual ccd color and monochrome camera, by Olympus (1 mentions)
8 protocols using cellsens image acquisition software
1
In Vitro Transcription and Microinjection of Fluorescent Reporter RNAs
2
Histological Analysis of Medaka Gill and Gallbladder
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Adult medaka were anesthetized using MS-222 at 12 weeks post-hatching and standard length and body weight were measured. Gills and gallbladders were removed and fixed in Bouin’s solution for 20 h, and then stored in 70% ethanol at 4 °C until needed. For histological analysis, the fixed organs were embedded in paraffin, cut into 6-µm-thick cross sections, and stained with hematoxylin and eosin. The sections were visualized using an Eclipse E600 microscope (Nikon, Tokyo, Japan). Images were acquired with a DP73 camera and cellSens image acquisition software (Olympus).
3
DLD1 Sphere Formation Assay
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DLD1 cells were cultured in DMEM/F-12 with epidermal growth factor (20 ng/mL), basic fibroblast growth factor (20 ng/mL), and B27 supplements in noncoated dishes. For serial passaging, the sphere cells were dissociated into single cells with Accutase (Invitrogen) once every 4–7 days and incubated. After the secondary generation, the DLD1 sphere cells were used for the experiments. The cells (5 × 104) were cultured in 6-well plates in the absence or presence of A3373 for 4 days. The cells were fixed with 4% paraformaldehyde, and sphere formation was calculated from 10 positions in each well by an Olympus BX53 microscope and cellSens image acquisition software (Olympus).
4
Embryonic Blood Cell Cytomorphology
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Peripheral blood cells from E9.5 to E12.5 embryos were collected as detailed above and cytospun onto glass slides at 400 g for 10 min (Cellspin® III, Tharmac). Slides were stained with Modified Wright’s Stain on a Siemens hematek 3000 system. The slides were viewed using an Olympus BX52 microscope (Olympus, Tokyo, Japan) with an Olympus UPlanApo 100x/1.35 NA Oil Iris objective (1,000 × total magnification). Micrographs were taken with an Olympus DP72 digital camera and with CellSens image acquisition software (Olympus, Tokyo, Japan).
5
Cardiac Tissue Preservation and Histological Analysis
6
Disc Morphology in Lumbar and Tail
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To provide an impression of disc gross morphological differences between lumbar and tail discs, individual discs were examined and photodocumented with an Olympus SZ61 stereomicroscope equipped with a digital SC30 camera and laptop running cellSens image acquisition software (all Olympus).
7
Histological Analysis of Aortic Tissue
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After euthanasia, the aorta was collected and washed with ice-cold sterile normal saline solution, fixed in 4% paraformaldehyde for 12 h at 4 °C. Tissues were dehydrated and cleared by graded ethanol and xylene, respectively, and embedded in paraffin. Paraffin blocks were cut into 5-µm thickness and stained with hematoxylin and eosin (H & E) for histological examination. Slides were visualized under a light microscope (Ni-U model eclipse; Nikon, Tokyo, Japan) with cellSens image acquisition software (Olympus, Tokyo, Japan).
8
Quantifying Serendipita Effects on Switchgrass
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To visualize and estimate the effect of Serendipita colonization on switchgrass root architecture, root hair density and surface area were measured at the precontact stage of colonization (Fig. 3). Images of the root tip were acquired using an Olympus SZX 12 fluorescent stereomicroscope (Olympus Corporation, Tokyo, Japan) equipped with an Olympus DP-80 dual CCD color and monochrome camera, and run by Olympus cellSens image acquisition software. Subsequently, a uniform fragment of 2 mm was selected from the root maturation zone for the estimation of root hair density and root surface area using RhizoVision Analyzer (version 1.0.3) (Seethepalli et al. 2019) (link). Within each colonization treatment, the data were analyzed by one-way analysis of variance using CoStat statistical software 6.4 (Cohort, Berkeley, CA, U.S.A.). Treatment means were compared using the least significant difference value at P < 0.05. The data were plotted graphically using the package ggplot2 (Wickham 2016) in R studio (R Studio, Inc., Boston, MA, U.S.A.).
Estimation of plant biomass.
Plants were harvested at late postcontact stage of colonization for the measurement of height, tiller diameter, and shoot dry biomass (Fig. 4). The data analysis and the graphical presentation follows the same procedure as for root hair density.
Estimation of plant biomass.
Plants were harvested at late postcontact stage of colonization for the measurement of height, tiller diameter, and shoot dry biomass (Fig. 4). The data analysis and the graphical presentation follows the same procedure as for root hair density.
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