Hearts were dissected from adult zebrafish as described (Ding et al., 2011 (link)). Following dissection, adult zebrafish hearts were placed on a microscope slide and imaged on a dissection microscope (Nikon) with top-lighting. Hearts were rotated to present maximal ventricular area and images were captured with Spot Imaging Software. Ventricular area was calculated with FIJI software (www.fiji.sc/ ). Prior to dissection, anesthetized zebrafish were partially dried with paper towels and weighed. Zebrafish mass was used as a normalizing factor for heart size index (VA/BW) since it is a greater variable than body length in adult fish.
Dissection microscope
Manufactured by Nikon
Sourced in Japan
The Nikon Dissection Microscope is a high-quality optical instrument designed for detailed observation and examination of specimens. It features a stable, ergonomic design and provides a clear and magnified view of the subject matter.
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Lab products found in correlation
Proparacaine hydrochloride, by Bausch & Lomb (2 mentions)
Rabbit serum, by Merck Group (1 mentions)
Rabbit anti asialo gm1, by Cedarlane (1 mentions)
Bx51 phase contrast microscope, by Olympus (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Quick rna tissue insect kit, by Zymo Research (1 mentions)
Igf 1, by RayBiotech (1 mentions)
Rankl, by RayBiotech (1 mentions)
Oct compound, by Thermo Fisher Scientific (1 mentions)
Igf 1, by R&D Systems (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Rankl, by R&D Systems (1 mentions)
Il 10, by R&D Systems (1 mentions)
Il 10, by RayBiotech (1 mentions)
Tnf α, by RayBiotech (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Mct20, by Scanco (1 mentions)
Mct 40 scanner, by Scanco (1 mentions)
A5040, by Merck Group (1 mentions)
16 protocols using dissection microscope
1
Zebrafish Heart Dissection and Imaging
2
Intraocular Delivery of Optogenetic Constructs
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Intraocular injections were performed as previously described by Wang et al.49 (link) Briefly, AAVs with optogenetic constructs were injected into the anterior chamber of the eye under ketamine anesthesia. As a local anesthetic, 1 drop of 0.5% proparacaine hydrochloride (Bausch & Lomb, Tampa, FL) was applied to each eye. To create a small aperture to the anterior chamber, the cornea was punctured using a sterile 33G needle (STERiJECT) under a dissection microscope (Nikon, Tokyo, Japan). After removing the needle, a fine borosilicate glass capillary (World Precision Instruments 1B150-4; Sarasota, FL) was connected to a Hamilton syringe using polyethylene tubing and passed through the same hole to introduce a small air bubble. The capillary was slowly removed and filled with 2-µL viral constructs (1010 infectious U/µL) diluted in Fast Green blue dye to monitor the injection through the puncture site. The capillary was then reinserted, and the anterior chamber filled with the distinct blue tint of the virus. The glass capillary was kept in position to monitor the pattern of the staining. To prevent leakage of the virus and to seal off the puncture site, the air bubble was gently moved to the insertion site. At the end of the procedure, neomycin was applied to decrease the risk of infection.
3
Metastatic B16F10 Melanoma Model
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Cultured B16F10 cells were recovered by Detachin (Gelantis) treatment and then injected into the tail veins of C57BL/6 mice (2 × 105 cells/200 μl DPBS or 5 × 105 cells/200 μl DPBS). After 14 day of tumor implantation, the mice were euthanized and the lungs were fixed in Fekete's solution. The number of nodules of metastatic tumor cells in the lungs was determined by using a dissection microscope (Nikon). To deplete the NK cells, mice were administered i.p. injections of 10 μL rabbit anti-asialo-GM1 (Cedarlane Laboratories Ltd.) while the control mice were injected with rabbit serum (Sigma-Aldrich). The injections of anti-asialo-GM1 or rabbit serum were performed 1 day before an i.v. injection of B16F10 cells, and then twice a week administration until euthanasia.
4
Copepod Morphological Examination Protocol
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Copepods were collected from the stagnant water retained in the burrows excavated by ocypodid crabs in two intertidal mud-flats using a hand net (0.2 mm mesh size) and also from near-bottom shallow waters using a light trap and a plankton net (0.2 mm mesh size) at high tide at dusk hours in eastern and southern Korea. For morphological examination, samples were fixed in 5% natural formalin-seawater solution. Specimens were later cleared in 70% lactic acid for 1 to 2 hours before dissecting under the dissection microscope (Nikon) in a drop of lactophenol on a wooden slide (Humes and Gooding 1964 ). The removed body parts and appendages were examined under a Olympus BX51 phase contrast microscope up to ×1,000. Drawings were made with the aid of a drawing tube attached to the microscope.
Body sizes of individuals were measured using a stage micrometer from the head to the tip of the caudal rami excluding caudal setae. The morphological terminology PageBreakfollows Huys and Boxshall (1991) . Abbreviations used in the text and figures are as follows: ae, aesthetasc; P1-P5, first to fifth swimming legs. Specimens are deposited at the National Institute of Biological Resources (NIBR), Incheon, Korea.
Body sizes of individuals were measured using a stage micrometer from the head to the tip of the caudal rami excluding caudal setae. The morphological terminology PageBreakfollows Huys and Boxshall (1991) . Abbreviations used in the text and figures are as follows: ae, aesthetasc; P1-P5, first to fifth swimming legs. Specimens are deposited at the National Institute of Biological Resources (NIBR), Incheon, Korea.
5
Quantifying Myocardial Infarct Size
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Heart tissues were frozen at −20°C for 10 min and cut into 4 slices transversely. One of the middle slices was then incubated at 37°C for 10 min with 1.5% TTC (triphenyltetrazolium chloride). Gross photographs were taken using a dissection microscope (Nikon). The images were analyzed by computerized planimetry using MetaVue image analysis software (Molecular Devices). The area of myocardial cross-section showing white color was defined as infarct, and the region in red was defined as the area at risk. Infarct size was expressed as a percentage of the area at risk.
6
Intraocular Viral Vector Injection
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Intraocular injections were performed according to Wang et al. (38 (link)). Briefly, AAV2-s or lentivirus with optogenetic constructs was injected into the anterior chamber of the eye of C57BL/6 mice anesthetized with ketamine (100 mg/kg) in ddH2O. In addition, one drop of 0.5% proparacaine hydrochloride (Bausch & Lomb, Tampa, FL) was applied to each eye as a local anesthetic. The cornea was punctured with a sterile 33-gauge needle (STERiJECT) under observation through a dissection microscope (Nikon) to create a small aperture to the anterior chamber. The needle was removed, and a fine borosilicate glass capillary (World Precision Instruments, 1B150-4) connected to a Hamilton syringe by polyethylene tubing was passed through the same hole to introduce a small air bubble. The capillary was slowly removed and filled with 2 μl of viral constructs (1010 infectious units/μl) diluted in Fast Green blue dye to facilitate and monitor the injection through the same puncture site. After introduction of the diluted virus, the anterior chamber was filled with a distinct blue tint, and the glass capillary was left in position for an additional minute to monitor the pattern of the staining. The air bubble was then gently moved to the insertion site to seal off the puncture and prevent leaking of vector. After the procedure, neomycin was applied to each eye to prevent infection.
7
Isolating Achilles Tendon Enthesis Tissue
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The site at which the P14 or P60 WT, Hyp, and C–/– Achilles tendon insert into the calcaneus was identified under a dissection microscope (Nikon) at ×5 magnification and isolated using a microsurgical knife by slicing through the proximal end of the tendon and the region adjacent to the distal portion of the calcaneus (14 (link)). The enthesis tissue was homogenized in Trizol (Thermo Scientific Fisher). Total RNA was precipitated using 100% alcohol, purified using the Quick-RNA Tissue/Insect Kit (Zymo Research).
8
Alveolar Bone Loss Analysis in Mice
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The maxillae of each mouse collected at the endpoint were defleshed and soaked in toluidine blue solution. Stained maxillae were photographed using a dissection microscope (Nikon), and total alveolar bone loss was calculated by measuring the cemento-enamel junction (CEJ) to the alveolar bone crest decalcification (ABC) distances on the buccal side of each root [41 (link)]. Some maxillae were fixed in 10% formaldehyde and then 10% EDTA solution at 4 °C for 3 to 4 weeks. The decalcified maxillae samples were then embedded in paraffin and sectioned (7-μm thickness) for TRAP and hematoxylin and eosin (H&E) staining. For immunofluorescence staining, the decalcified maxillae samples were embedded in OCT compound (Fisher Scientific) and sectioned (6 μm thickness) using a cryostat. The expression of Sema4D was monitored using biotin-conjugated anti-Sema4D-mAb, followed by avidin conjugated to AlexaFluor 488. Gingival tissue homogenates were obtained from the gingival tissue of each mouse at the endpoint, and Sema4D, TNF-α, RANKL, OPG, IL-10 and IGF-1 production in the homogenate was analyzed using an ELISA kit purchased from the following sources: Sema4D: RayBiotech; TNF-α, RANKL, OPG, IL-10 and IGF-1: R&D Systems.
9
Tissue Imaging with Nikon Microscope
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After dissection, tissues were imaged under a Nikon dissection microscope using NIS-Elements software. Either bright field or epifluorescence of either red or green channels were used for imaging.
10
Ear and Skin Organoid Injury
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For ear wounding, we used a standard 2mm mechanical punch (Roboz, Gaithersburg, MD) to create a hole in the center of each outer ear (pinna). Ear hole diameter was measured using a dissection microscope (Nikon) in the horizontal and vertical directions on a weekly basis. Ears were excluded if there were signs of wound infection, tearing of the ear, or abnormal geometric shape. These criteria were pre-established.
For human skin organoid injury, we used a standard 1.5mm punch (Acupunch) to create a hole in the center of each graft.
For human skin organoid injury, we used a standard 1.5mm punch (Acupunch) to create a hole in the center of each graft.
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