Imaging:Article Title: The ROCK1 PHC1 domain mediates recruitment to Rho activity zones and contractility signaling.
Article Snippet: TIRF microscopy was performed on an Olympus IX-81 microscope, equipped with an Apo TIRF 60 × 1.45 NA oil immersion objective, a TIR-MITICO–motorized TIRF illumination combiner, and a ZDC autofocus device. .. Four different lasers were employed for imaging cells: a 514-nm OBIS diode laser (150 mW) (Coherent, Inc.) and Cell R diode lasers (Olympus) with wavelengths of 405 nm (50 mW), 445 nm (50 mW), and 561 nm (100 mW). .. In addition, wide-field illumination was performed using the Spectra X light engine (Lumencor).
Concentration Assay:Article Title: Protocol for quantifying chromatin DNA replication by multiplex labeling and single-molecule localization microscopy.
Article Snippet: .. 405 nm laser Coherent OBIS 561 nm laser MPB Communications 2RU-VFL-P-1000-561-B1R 647 nm laser MPB Communications 2RU-VFL-P-1000-647-B1R Near-infrared focus-lock laser Thorlabs LP940-SF30 Preparation of secondary antibody stock Reagent Stock concentration Storage Final concentration Volume Secondary antibody 1.0–1.3 mg/mL 4◦C 0.5–0.6 mg/mL 1 mL Glycerol 80% − 20◦C 40% 1 mL NaN3 10% − 20 ◦C 0.05% 10 μL Cy3B labeling reaction recipe Reagent Stock concentration Storage Final concentration Volume Secondary antibody stock ∼0.5 mg/mL, ∼3.3 μM − 20◦C 0.25 mg/mL, ∼1.7 μM ∼100 μL NaHCO3 1 M, freshly prepared − 20 ◦C 0.1 M 20 μL Cy3B-NHS dye 500–1000 μM − 20◦C 50–100 μM 20 μL PBS 13 20◦C–25◦C 13 up to 200 μL Reagents for cell-cycle synchronization and DNA labeling Reagent Stock concentration Working concentration Solvent Thymidine 200 mM (1003) 2 mM sterile water Aphidicolin 200 μg/mL (1003) 2 μg/mL DMSO EdU 10 mM (10003) 10 μM DMSO BrdU 16 mM (10003) 16 μM DMSO 6 STAR Protocols 7, 104882, December 18, 2026 Prepare thymidine, aphidicolin, EdU, and BrdU stock solutions in advance, aliquot them to avoid repeated freeze-thaw cycles, and prepare fresh working solutions in pre-warmed complete DMEM on the day of use. ..
Labeling:Article Title: Protocol for quantifying chromatin DNA replication by multiplex labeling and single-molecule localization microscopy.
Article Snippet: .. 405 nm laser Coherent OBIS 561 nm laser MPB Communications 2RU-VFL-P-1000-561-B1R 647 nm laser MPB Communications 2RU-VFL-P-1000-647-B1R Near-infrared focus-lock laser Thorlabs LP940-SF30 Preparation of secondary antibody stock Reagent Stock concentration Storage Final concentration Volume Secondary antibody 1.0–1.3 mg/mL 4◦C 0.5–0.6 mg/mL 1 mL Glycerol 80% − 20◦C 40% 1 mL NaN3 10% − 20 ◦C 0.05% 10 μL Cy3B labeling reaction recipe Reagent Stock concentration Storage Final concentration Volume Secondary antibody stock ∼0.5 mg/mL, ∼3.3 μM − 20◦C 0.25 mg/mL, ∼1.7 μM ∼100 μL NaHCO3 1 M, freshly prepared − 20 ◦C 0.1 M 20 μL Cy3B-NHS dye 500–1000 μM − 20◦C 50–100 μM 20 μL PBS 13 20◦C–25◦C 13 up to 200 μL Reagents for cell-cycle synchronization and DNA labeling Reagent Stock concentration Working concentration Solvent Thymidine 200 mM (1003) 2 mM sterile water Aphidicolin 200 μg/mL (1003) 2 μg/mL DMSO EdU 10 mM (10003) 10 μM DMSO BrdU 16 mM (10003) 16 μM DMSO 6 STAR Protocols 7, 104882, December 18, 2026 Prepare thymidine, aphidicolin, EdU, and BrdU stock solutions in advance, aliquot them to avoid repeated freeze-thaw cycles, and prepare fresh working solutions in pre-warmed complete DMEM on the day of use. ..
Article Title: Ligand-dependent G protein dynamics underlying opioid efficacy.
Article Snippet: .. Gαi-Δ6-90/238C labeled with LD555 (donor) and JFX673-peg4 (acceptor) was excited using a 532-nm solid-state laser (Coherent OBIS LS laser) set to 40 mW. .. Photons emitted from LD555 and JFX673 were collected through a ×60 (numerical aperture: 1.27) water-immersion objective (Nikon) and split onto two cooled electron-multiplying charge-coupled device cameras (Andor iXon Ultra 888) connected by a 640-nm dichroic mirror (ZT640rdc, Chroma) housed in a TwinCam (Cairn).
DNA Labeling:Article Title: Protocol for quantifying chromatin DNA replication by multiplex labeling and single-molecule localization microscopy.
Article Snippet: .. 405 nm laser Coherent OBIS 561 nm laser MPB Communications 2RU-VFL-P-1000-561-B1R 647 nm laser MPB Communications 2RU-VFL-P-1000-647-B1R Near-infrared focus-lock laser Thorlabs LP940-SF30 Preparation of secondary antibody stock Reagent Stock concentration Storage Final concentration Volume Secondary antibody 1.0–1.3 mg/mL 4◦C 0.5–0.6 mg/mL 1 mL Glycerol 80% − 20◦C 40% 1 mL NaN3 10% − 20 ◦C 0.05% 10 μL Cy3B labeling reaction recipe Reagent Stock concentration Storage Final concentration Volume Secondary antibody stock ∼0.5 mg/mL, ∼3.3 μM − 20◦C 0.25 mg/mL, ∼1.7 μM ∼100 μL NaHCO3 1 M, freshly prepared − 20 ◦C 0.1 M 20 μL Cy3B-NHS dye 500–1000 μM − 20◦C 50–100 μM 20 μL PBS 13 20◦C–25◦C 13 up to 200 μL Reagents for cell-cycle synchronization and DNA labeling Reagent Stock concentration Working concentration Solvent Thymidine 200 mM (1003) 2 mM sterile water Aphidicolin 200 μg/mL (1003) 2 μg/mL DMSO EdU 10 mM (10003) 10 μM DMSO BrdU 16 mM (10003) 16 μM DMSO 6 STAR Protocols 7, 104882, December 18, 2026 Prepare thymidine, aphidicolin, EdU, and BrdU stock solutions in advance, aliquot them to avoid repeated freeze-thaw cycles, and prepare fresh working solutions in pre-warmed complete DMEM on the day of use. ..
Solvent:Article Title: Protocol for quantifying chromatin DNA replication by multiplex labeling and single-molecule localization microscopy.
Article Snippet: .. 405 nm laser Coherent OBIS 561 nm laser MPB Communications 2RU-VFL-P-1000-561-B1R 647 nm laser MPB Communications 2RU-VFL-P-1000-647-B1R Near-infrared focus-lock laser Thorlabs LP940-SF30 Preparation of secondary antibody stock Reagent Stock concentration Storage Final concentration Volume Secondary antibody 1.0–1.3 mg/mL 4◦C 0.5–0.6 mg/mL 1 mL Glycerol 80% − 20◦C 40% 1 mL NaN3 10% − 20 ◦C 0.05% 10 μL Cy3B labeling reaction recipe Reagent Stock concentration Storage Final concentration Volume Secondary antibody stock ∼0.5 mg/mL, ∼3.3 μM − 20◦C 0.25 mg/mL, ∼1.7 μM ∼100 μL NaHCO3 1 M, freshly prepared − 20 ◦C 0.1 M 20 μL Cy3B-NHS dye 500–1000 μM − 20◦C 50–100 μM 20 μL PBS 13 20◦C–25◦C 13 up to 200 μL Reagents for cell-cycle synchronization and DNA labeling Reagent Stock concentration Working concentration Solvent Thymidine 200 mM (1003) 2 mM sterile water Aphidicolin 200 μg/mL (1003) 2 μg/mL DMSO EdU 10 mM (10003) 10 μM DMSO BrdU 16 mM (10003) 16 μM DMSO 6 STAR Protocols 7, 104882, December 18, 2026 Prepare thymidine, aphidicolin, EdU, and BrdU stock solutions in advance, aliquot them to avoid repeated freeze-thaw cycles, and prepare fresh working solutions in pre-warmed complete DMEM on the day of use. ..
Sterility:Article Title: Protocol for quantifying chromatin DNA replication by multiplex labeling and single-molecule localization microscopy.
Article Snippet: .. 405 nm laser Coherent OBIS 561 nm laser MPB Communications 2RU-VFL-P-1000-561-B1R 647 nm laser MPB Communications 2RU-VFL-P-1000-647-B1R Near-infrared focus-lock laser Thorlabs LP940-SF30 Preparation of secondary antibody stock Reagent Stock concentration Storage Final concentration Volume Secondary antibody 1.0–1.3 mg/mL 4◦C 0.5–0.6 mg/mL 1 mL Glycerol 80% − 20◦C 40% 1 mL NaN3 10% − 20 ◦C 0.05% 10 μL Cy3B labeling reaction recipe Reagent Stock concentration Storage Final concentration Volume Secondary antibody stock ∼0.5 mg/mL, ∼3.3 μM − 20◦C 0.25 mg/mL, ∼1.7 μM ∼100 μL NaHCO3 1 M, freshly prepared − 20 ◦C 0.1 M 20 μL Cy3B-NHS dye 500–1000 μM − 20◦C 50–100 μM 20 μL PBS 13 20◦C–25◦C 13 up to 200 μL Reagents for cell-cycle synchronization and DNA labeling Reagent Stock concentration Working concentration Solvent Thymidine 200 mM (1003) 2 mM sterile water Aphidicolin 200 μg/mL (1003) 2 μg/mL DMSO EdU 10 mM (10003) 10 μM DMSO BrdU 16 mM (10003) 16 μM DMSO 6 STAR Protocols 7, 104882, December 18, 2026 Prepare thymidine, aphidicolin, EdU, and BrdU stock solutions in advance, aliquot them to avoid repeated freeze-thaw cycles, and prepare fresh working solutions in pre-warmed complete DMEM on the day of use. ..
Fluorescence:Article Title: Localization and Trafficking Behavior of SPPL2A During Mouse Spermiogenesis.
Article Snippet: Brightfield and epiflu‐ orescence imaging were performed on an inverted microscope (IX83, Evident Scientific, Inc., Tokyo, Japan) equipped with a 100×, 1.49 NA objective and a 2× magnification lens. .. Fluorescence illumination was performed with a free‐space 15‐mW, 561‐nm laser (OBIS, Coherent Technology Corp., Saxonburg, PA, USA), cleaned by a bandpass filter (ZET405/488/561/647X, Chroma Technology Corp., Bellows Falls, VT, USA), reflected by a quad‐band dichroic mirror (ZT405/488/561/647RPC, Chroma Technology Corp.), and dis‐ tributed across a 2000 μm2 sample area. .. Emission passed through a 525/50M emission filter (Evident Scientific, Inc.) and a 4‐band notch filter (ZET405/488/561/640m; Chroma Technology Corp.) before being detected by an EMCCD camera (iXon 897‐Ultra, Andor Technology Ltd.) with 150 EM gain, 20 ms exposure, and 47 Hz frame rate.
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