After the daily administration of vehicle or CDN1163 (40 mg/kg) for 1 week, muscular damage was induced by a modified treadmill protocol as previously described (26 ). Briefly, the exercise load began at a speed of 5 m/min for 5 min and gradually increased by 1 m/min every minute until the speed reached 10 m/min. The mice were kept on the treadmill for 30 min under repeated gentle nudges. After a 10-min interval, the session was repeated. All mice were able to carry out the test until the end of the experiment. After treadmill exercise, the mice were injected intraperitoneally with EBD (10 mg/ml, 0.1 ml/10 g BW). The next day, the mice were sacrificed, and their GC muscles were cut into cryosections to observe EBD uptake (red fluorescence) by degenerating fibers. Images were taken with a BZ-X810 fluorescence microscope (Keyence, Osaka, Japan).
Bz x810 fluorescence microscope
Manufactured by Keyence
Sourced in Japan
The BZ-X810 fluorescence microscope is a tool designed for high-resolution imaging and analysis of fluorescently-labeled samples. It features a compact and modular design, allowing for versatile configuration to suit various research and diagnostic needs. The core function of the BZ-X810 is to capture and process fluorescence images, enabling detailed examination of cellular structures, protein localization, and other fluorescent-based applications.
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Lab products found in correlation
70 m cell strainer, by BD (4 mentions)
Bz x810 analyzer software, by Keyence (4 mentions)
Costar ultra low attachment plate, by Corning (3 mentions)
Stempro accutase, by Thermo Fisher Scientific (3 mentions)
Psc cardiomyocyte differentiation kit, by Thermo Fisher Scientific (2 mentions)
Human three germ layer 3 color immunocytochemistry kit, by R&D Systems (2 mentions)
Vectashield antifade mounting media with dapi, by Vector Laboratories (2 mentions)
Paraformaldehyde (pfa), by Fujifilm (2 mentions)
Click it edu imaging kit, by Thermo Fisher Scientific (2 mentions)
Dio lipophilic dye, by Thermo Fisher Scientific (2 mentions)
Cytopainter red, by Abcam (2 mentions)
Tissue tek oct compound, by Sakura Finetek (2 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (2 mentions)
Geltrex, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Sytox green nucleic acid stain, by Thermo Fisher Scientific (1 mentions)
Cls3603, by Corning (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Phalloidin ifluor 488, by Abcam (1 mentions)
D1306, by Thermo Fisher Scientific (1 mentions)
66 protocols using bz x810 fluorescence microscope
1
Muscular Damage Evaluation via Treadmill
2
Orthotopic GBM Xenograft Model
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Orthotopic xenograft model of GSC-derived GBM was generated by transplantation of 5 × 10 4 TGS-01 GSCs into the brain of 4-week-old female nude mice (BALB/cSlc-nu/nu, SLC, Shizuoka, Japan). Briefly, a small burr hole was drilled in the skull 0.5 mm anterior and 2.0 mm lateral from bregma with a micro drill, and dissociated cells were transplanted at a depth of 3 mm below the dura mater. Mice were sacrificed at the indicated time points or upon occurrence of neurological symptoms. Mouse brains were fixed with 4% paraformaldehyde solution, embedded in paraffin, and then sectioned at a thickness of 5 μm. Sections were stained with Hematoxylin and Eosin (H&E). The sections were captured using a BZ-X810 fluorescence microscope (Keyence). All animal experiments were approved by the Committees on Animal Experimentation of Gifu Pharmaceutical University and Kanazawa University and performed in accordance with the guidelines for the care and use of laboratory animals. The numbers of animals used per experiment are stated in the figure legends.
3
Orthotopic GBM Xenograft Model
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Orthotopic xenograft model of GSC-derived GBM was generated by transplantation of 5 × 10 4 TGS-01 GSCs into the brain of 4-week-old female nude mice (BALB/cSlc-nu/nu, SLC, Shizuoka, Japan). Briefly, a small burr hole was drilled in the skull 0.5 mm anterior and 2.0 mm lateral from bregma with a micro drill, and dissociated cells were transplanted at a depth of 3 mm below the dura mater. Mice were sacrificed at the indicated time points or upon occurrence of neurological symptoms. Mouse brains were fixed with 4% paraformaldehyde solution, embedded in paraffin, and then sectioned at a thickness of 5 μm. Sections were stained with Hematoxylin and Eosin (H&E). The sections were captured using a BZ-X810 fluorescence microscope (Keyence). All animal experiments were approved by the Committees on Animal Experimentation of Gifu Pharmaceutical University and Kanazawa University and performed in accordance with the guidelines for the care and use of laboratory animals. The numbers of animals used per experiment are stated in the figure legends.
4
Immunofluorescence Staining of HRTV N Protein
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Virus-infected cells were washed twice with PBS and fixed with 4% paraformaldehyde (PFA) in PBS for 30 min at room temperature. Next, the cells were permeablized with 0.2% Triton X-100 for 1 h and washed three times with PBS. After blocking with PBS- 3% BSA for 1 hour, cells were incubated with mouse monoclonal antibody (2AG8) against the HRTV N protein at 4°C overnight. After three additional washes in PBS, cells were incubated with the goat anti-mouse secondary antibody conjugated with Alexa Fluor 594 (Invitrogen #A-11005) for 1 h at room temperature. The nuclei were stained with DAPI for 2 min and then washed with PBS for 3 times. Images were collected using Keyence BZ-X 810 fluorescence microscope.
5
Hematoxylin and Eosin Staining of Pancreatic Tissue
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Deparaffinized pancreatic sections were stained with hematoxylin and eosin by the conventional method. The slides were mounted in malinol (Muto). Images were taken using a BZ-X810 fluorescence microscope (Keyence).
6
NTM Cell Staining Protocol
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After enzymatic treatment of 1E5 NTM cells, 50 µL of the reaction was mixed with 50 µL of a stain mix containing 10 mM of SYTOX Green Nucleic Acid Stain (SG) (Invitrogen) and 0.25 mM FM4-64 (FM) (Invitrogen) in PBS in a black 96-well plate (Corning CLS3603). The plate was centrifuged for 2 min at 3,000 rpm and imaged with a Keyence BZ-X810 Fluorescence Microscope at 40× magnification. Unprocessed images were overlaid and resized using Photoshop 2022.
7
Immunofluorescence Analysis of Osteoblast and Osteocyte Markers
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Transwell insert membranes, seeded with OBs and OCy-containing collagen gels were removed from the inserts. Remaining 24 -well plates with PBMC/differentiated OCs as well as other samples were washed once with PBS and cells were fixed by incubation with phosphate buffered formaldehyde (4 %) for at least 1 h. After permeabilisation (0.1 % Triton X-100 in PBS for 5 min) and blocking (1 % BSA in PBS/30 min) OBs and OCs were incubated with DAPI (ThermoFisher Scientific) to detect cell nuclei and Phalloidin-iFluor 488 (Abcam) to visualise cytoskeleton (1 h protected from light). To observe specific biomarkers and morphological changes, OCys were stained for dentin matrix protein 1 (DMP1) and sclerostin (SOST). Primary and secondary antibodies were diluted in 1 % BSA in PBS. Rabbit anti-human DMP1 (TaKaRa; dilution 1:600; working concentration: 3.3 μg/mL) respectively rabbit anti-human SOST (antibodies-online GmbH; dilution 1:100; working concentration: 100 μg/mL) was detected with AlexaFluor 546-conjugated goat anti-rabbit secondary antibody (Abcam; 1:250). Secondary antibodies were added together with DAPI and Phalloidin-iFluor 488. Z-stack images were acquired using a Keyence BZ-X810 fluorescence microscope.
8
Immunofluorescence Staining for PAX8 and Ki67
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Tissues and organoids were fixed in 4% paraformaldehyde (PFA) (BL539A, Biosharp) for 30 min, dehydrated with sucrose, and embedded in 7.5% gelatin for standard histology and immunofluorescence (IF). Frozen sections (10 mm) of the embedded samples were retrieved using citric acid (PH6.0). The adherent cells were fixed using 4% PFA for 20 min. Both adherent cells and frozen sections were permeabilized using 0.25% Triton X-100 in PBST, and blocked with Primary Antibody Dilution Buffer (E674004, Sangon Biotech, China). After overnight incubation with 1:1000 primary antibody Rabbit anti-PAX8 (10336-1-AP, Proteintech) and anti-Ki67 (MA5-14520, ThermoFisher) at 4°C, sections were washed two times in 0.125% PBST and incubated at room temperature with 1:1000 secondary antibody anti-rabbit (Cy3) (711-165-152, Jackson). Then, they were incubated with 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI) (D1306, ThermoFisher) stain solution at room temperature. After washing three times with 0.125% PBST, they were scanned using a Keyence BZ-X810 Fluorescence Microscope and the proportions of immunoreactive cells were counted. For H&E staining, the sections were washed twice and stained using a hematoxylin-eosin (HE) Stain Kit (G1120, Solarbio, China) according to the manufacturer’s instructions.
9
Pluripotency and Trilineage Immunofluorescence
10
Immunohistochemical Analysis of Pancreatic Insulin and CD45
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Pancreatic sections were deparaffinized, blocked, and loaded with anti-insulin (Cell Signaling, 1:400) or anti-CD45 (1:200) antibodies as described in section 2.10 . The slides were loaded with anti-rabbit POD conjugate (TaKaRa) and incubated for 30 min at room temperature. The slides were then loaded with DAB substrate solution (Nichirei) and incubated for 30 min at room temperature. The slides were washed with water, counterstained with hematoxylin, dehydrated, and mounted in malinol (Muto). Images were taken using a BZ-X810 fluorescence microscope (Keyence).
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