We anesthetized zebrafish from 2 to 6 dpf with 0.0168% tricaine (Sigma-Aldrich, catalog no. MS-222, E1052) and acquired images in brightfield using the Zeiss stereomicroscope at 5× (Fig. 4A ) and 10× (Fig. 4, B and C ). We recorded videos of the hearts beating for 30 s per zebrafish with the Zeiss stereomicroscope at 10× (movies S1 to S9). Beats were counted for 30× and doubled to achieve beats per minute.
Stereomicroscope
Manufactured by Zeiss
Sourced in Germany, United States, Japan
The Zeiss Stereomicroscope is a precision optical instrument designed for high-resolution, three-dimensional observation and analysis. It provides a magnified, stereoscopic view of samples, enabling users to examine specimens in detail. The core function of the Stereomicroscope is to deliver a clear, binocular image of the subject matter.
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Lab products found in correlation
Floxal, by Bausch & Lomb (6 mentions)
Conjuncain, by Bausch & Lomb (5 mentions)
Xylazine, by Bayer (4 mentions)
Ketamine, by Ratiopharm (3 mentions)
Axiocam erc5s camera, by Zeiss (3 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Ds 11, by DeNovix (2 mentions)
M2 medium, by Merck Group (2 mentions)
Formaldehyde, by Avantor (2 mentions)
Crystal violet, by Merck Group (2 mentions)
William s e medium, by Thermo Fisher Scientific (2 mentions)
Griener 90 well plate, by Merck Group (2 mentions)
24 well plate, by Thermo Fisher Scientific (2 mentions)
Zen software, by Zeiss (2 mentions)
Axiovision software, by Zeiss (2 mentions)
Axio zoom v16, by Zeiss (2 mentions)
Axio imager m2, by Zeiss (2 mentions)
Human chorionic gonadotropin (hcg), by Merck Group (2 mentions)
Hyaluronidase, by Merck Group (2 mentions)
Lsm 780, by Zeiss (2 mentions)
183 protocols using stereomicroscope
1
Zebrafish Cardiac Imaging and Analysis
2
Thymus Organ Culture Protocol
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Mice were sacrificed for thymus collection. After the surface connective tissue was removed, the thymus was washed three times with PBS containing 1% penicillin-streptomycin solution and placed into cell supernatants collected from 2.5.1. All of the thymus were cultured in a constant temperature incubator at 37 °C with 5% CO2 and saturated humidity for 7 days. The morphological changes of thymus organs were recorded using a stereo microscope (Zeiss, Jena, Germany).
3
Embryonic Stem Cell Characterization
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Mouse embryos (E7.75) were dissected using forceps under a stereomicroscope (Zeiss) and regions of interest were dissociated and harvested using TrypLE. Embryoid bodies (EBs) and cells were dissociated and harvested using TrypLE. Single-cells were analyzed for RFP/GFP expression or sorted using a SH800 Cell sorter (Sony Biotechnologies). Live cells were analyzed for RFP and GFP expression and stained with antibodies targeting for the presence of appropriate markers. Cells were stained with the following antibodies: anti-mouse Cxcr4 conjugated with PerCP-eFluor 710 (1:200; 46-9991-80 eBiosciences) anti-mouse EphA2 conjugated with APC (1:100; Cat. FAB639A R&D systems), anti-human Cxcr4 conjugated with PE or APC (1:25; Cat. FAB170P R&D systems). For cTNT and Isl1 expression, cells were fixed with 4% paraformaldehyde (PFA) for 10 min, permeabilized with saponin (Sigma), stained with either mouse cTNT (1:500, Cat. MS-295-P1 Thermo Scientific) or mouse Islet1 antibody (1:200, Cat. 39.3F7 Developmental Studies Hybridoma Bank, Iowa City, IA), followed by incubation with secondary antibody conjugated with Alexa Fluor 647 (1:500, Invitrogen). Data were analyzed using FlowJo software.
4
Ovary Anatomy and Reproductive Analysis
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Ovaries of mated female BPH and fruit flies were anatomized under a stereomicroscope (Zeiss). The ovaries were placed on a glass slide, and 50 μl PBS was added to float the ovaries to maintain their original shape. The ovary images were obtained by a light microscope with a digital video camera (Zeiss, ProgRes 3008 mF, Jenoptik, Jena, Germany) and were used to measure the area of individual ovaries and the number of eggs per ovary using the matching NIS-Elements software.
5
Touch-Evoked Scratch Reflex Assay
6
Analyzing Zebrafish Motor Behavior
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At 48 hpf, zebrafish embryos were tested for motor behavior under a stereomicroscope (Zeiss, Germany) using the touch-evoked escape response (TEER) experiment. Morphologically normal zebrafish from each experimental condition were touched lightly at the level of the tail with a tip, and their responses to the stimulus were recorded with a Grasshopper 2 Camera (Point Grey Research) at 30 Hz. The videos were then analyzed using the Manual Tracking plugin of the ImageJ software, and the swim duration, swim distance and maximum swim velocity of each embryo were calculated as previously described [49 (link)].
7
Colony-Forming Unit Assay for ASC Quantification
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To assess the percentage of colony-forming cells present in the SVF, a colony-forming-unit assay was performed, which is an assay to determine the percentage of ASC present in the SVF (Bourin et al. 2013 (link)). For this, SVF cells were seeded at a density of 10 and 100 cells/cm2 (in triplicate) and cultured in normal proliferation medium containing DMEM medium, supplemented with 10 % fetal bovine serum (FBS), 1 % penicillin/streptomycin and 1 % L-glutamine (all from Gibco). Cells were maintained at 37 °C, 95 % humidity and 5 % CO2. After 14 days, the cells were washed with PBS, fixed with 4 % formalin for 10 min, and subsequently stained in a 1 % toluidine blue solution in borax buffer for 1 min. After washing with water, colonies of approximately 50 cells or more were scored using a stereomicroscope (Zeiss, Germany).
8
Targeted Genome Editing in Axolotl
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For lineage tracing experiments, we targeted one site in an egfp transgene and endogenous tyrosinase gene, and two sites in a mCherry transgene and endogenous methyltransferase-like gene listed in Supplementary file 1 . Target gRNA sequences are listed in Supplementary file 1 . Axolotl matings and microinjections were carried out as previously described (Flowers and Crews, 2015 (link)), using 500 pg of sgRNA and 1000 pg of cas9 mRNA. cagg:egfp and cagg:nuclear mcherry transgenic animals were mated with non-transgenic animals. Successful mutagenesis of egfp and nuclear mcherry transgenic animals was confirmed by imaging animals using fluorescence and a Zeiss stereomicroscope. Successful mutagenesis of tyrosinase, meth t1, and meth t2 were confirmed using fragment analysis of fluorescent PCR products as previously described (Flowers and Crews, 2015 (link)).
To assess allelic diversity, matings and microinjections were carried out as before but 5 pg of each of 5 sgRNAs and either 100 or 1000 pg of cas9 mRNA were injected into each blastomere at the two-cell stage. At two weeks post-injection, 5 larvae from each injection condition were used for genomic DNA extraction, target amplification, and NGS.
To assess allelic diversity, matings and microinjections were carried out as before but 5 pg of each of 5 sgRNAs and either 100 or 1000 pg of cas9 mRNA were injected into each blastomere at the two-cell stage. At two weeks post-injection, 5 larvae from each injection condition were used for genomic DNA extraction, target amplification, and NGS.
9
Aedes aegypti Salivary Gland Extraction
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Aedes aegypti (Liverpool strain) mosquitoes were reared under standard conditions in a 12-h light/dark cycle at and maintained at 28 °C, 80% humidity, at the Laboratory of Malaria and Vector Research insectary, NIAID, NIH. Salivary glands from sugar-fed 5- to 8-day-old adult female mosquitoes were dissected in ice-cold PBS pH 7.4 using a stereomicroscope (Zeiss, Thornwood, NY, USA). Next, salivary gland extracts (SGE) was obtained by sonicating dissected salivary glands in PBS, pH 7.4 using a Branson Sonifier 450 (Branson Ultrasonics, Danbury, CT, USA). Supernatants were recovered after disrupted tissues were centrifuged for 5 min at 12,000× g. The concentration of protein content was measured by spectrophotometry at A280 (DS-11, DeNovix, Wilmington, DE, USA), then the SGE was kept at −80 °C until use.
10
Preparation of Bee Worker Tissue
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Tissue samples of B. ignitus worker bees were collected using a stereo microscope (Zeiss, Jena, Germany) and washed with phosphate-buffered saline (PBS; 140 mM NaCl, 27 mM KCl, 8 mM Na2HPO4, 1.5 mM KH2PO4; pH 7.4). The fresh tissues were used for total RNA and protein sample preparation.
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