4T1 cancer cells were incubated in 6-well plates under hypoxic condition at 37 °C for 24 h. After which, previous medium was replaced by fresh medium that contained free Ce6, MCC, and OxgeMCC-r (Ce6 concentration is 8 ppm) for 4 h co-culture. Thereafter, the cells were treated with a 671 nm laser irradiation (100 mW cm−2, 30 s). After co-culture for another 4 h, cells were stained with calcein AM (5 μL) and PI (10 μL) according to the product description for 0.5 h and then observed using a fluorescence inverted microscope (Olympus UHGLGPS, China).
U hglgps
Manufactured by Olympus
Sourced in Japan, China
The U-HGLGPS is a high-intensity mercury vapor lamp designed for use in microscopy and other laboratory applications. It provides a powerful, broad-spectrum light source for illumination and analysis purposes. The lamp's core function is to generate intense ultraviolet, visible, and near-infrared radiation to enable various microscopy techniques.
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Lab products found in correlation
Orca flash 4.0 cmos camera, by Hamamatsu Photonics (4 mentions)
Fvmpe rs, by Olympus (3 mentions)
Metamorph, by Molecular Devices (3 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Beyotime (3 mentions)
Tcs sp8 sted, by Leica (3 mentions)
Paraformaldehyde (pfa), by Biosharp (3 mentions)
Smz2000, by Nikon (2 mentions)
Anti dsrna antibody, by Techcomp Instruments (2 mentions)
Smz2000 microscope, by Nikon (2 mentions)
Orca r2, by Hamamatsu Photonics (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Lsm 510, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Triton x 100, by Solarbio (2 mentions)
Szx16, by Olympus (1 mentions)
Ab222783, by Abcam (1 mentions)
A1374, by ABclonal (1 mentions)
Cf488 tunel cell apoptosis detection kit, by Wuhan Servicebio Technology (1 mentions)
Gb21303, by Wuhan Servicebio Technology (1 mentions)
Bs 2072r, by Bioss Antibodies (1 mentions)
69 protocols using u hglgps
1
Photodynamic Therapy for 4T1 Cells
2
Fluorescence Imaging with Stereoscopic Microscopy
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A stereoscopic fluorescence microscope (SZX-16; Olympus) was used to acquire fluorescence images. A mercury lamp (U-HGLGPS; Olympus) was employed as light source for fluorescence excitation. The wavelength of excitation light was set at 405 nm using an optical filter (D405/20x; Chroma). Fluorescence from the sample was collimated with an objective lens (SDFPLAPO × 0.5; Olympus), transmitted through a long-pass filter (HQ430LP; Chroma) at wavelength longer than 430 nm, and imaged on a color charge-coupled device (CCD) camera (DP73; Olympus) with the exposure time of 1 s.
3
Immunofluorescence Staining for Cellular Markers
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Immunofluorescence staining was performed as previously described (Chen et al., 2020 (link)). In short, the frozen sections were rewarmed at room temperature. After antigen retrieval in citric acid antigen repair solution, the sections were incubated at 4°C overnight with the following primary antibodies: anti-CD31 (1:200, ab222783, Abcam, USA), anti-IBA-1 (1:400, 019-19741, FUJIFILM Wako Shibayagi, Japan), anti-CD68 (1:200, 28058-1-AP, Proteintech, China), anti-GFAP (1:400, #80788, CST, USA), anti-MPO (1:50, A1374, Abclonal, China), and anti-NeuN (1:200, #94403, CST, USA). Then, the sections were incubated with the appropriate fluorescently labeled secondary antibody at room temperature for 1 h. For double staining with TUNEL fluorescence, the procedure followed the instructions of the CF488 TUNEL Cell Apoptosis Detection Kit (G1504, Servicebio, Wuhan, China). After incubation with secondary antibody, the sections were equilibrated with Equilibrium Buffer for 10 min and then incubated at 37°C for 1 h with TDT incubation buffer. Finally, nuclei were stained with DAPI. The sections were observed and photographed with a fluorescence microscope (U-HGLGPS, Olympus, Japan). Micrographs were analyzed with ImageJ software (ImageJ, RRID: SCR _ 003070).
4
Immunofluorescence Staining of AQP5, NOS2, and ET-1
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Immunofluorescence staining was accomplished by deparaffinizing the above paraffin sections, repairing the antigen, and blocking them by drip for 30 minutes. Phosphate buffer solution (PBS) was then diluted the primary antibody before it was incubated at 4°C overnight. Fluorescein-labeled secondary antibodies were then incubated for 50 minutes at room temperature after washing. The sections were incubated for 10 minutes at room temperature in the darkness before the nuclei were counterstained with DAPI. After adding the autofluorescence quench agent for 5 minutes, the slides were then rinsed with running water for 10 minutes. The mounted slides were observed and analyzed using a fluorescence microscope (U-HGLGPS, OLYMPUS, Japan). Aquaporin 5 (AQP5) (NBP2-92926, NOVUS), Inducible nitric oxide synthase (NOS2) (bs-2072R, Bioss), and Endothelin-1 (ET-1) (NB300-526, NOVUS) were labeled by CY3 (Servicebio, GB21303; Servicebio, GB21301) as red fluorescence.
5
Trigeminal Nerve Imaging Preparation
6
Histological Analysis of Small Intestine
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After the procedure, samples of the small intestine were removed from all quadrants and embedded in paraffin prior to standard hematoxylin and eosin (H&E) staining. Analyses were performed using light microscopy (Olympus life Sciences U-HGLGPS).
7
Single-Cell Gel Electrophoresis for DNA Damage
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DNA damage was assessed using a single-cell gel electrophoresis assay under neutral conditions with a DNA Damage Detection Kit (SCGE) (KeyGEN BioTECH, KGA240) according to the manufacturer’s protocol. Briefly, the normal melting point agarose (NMA) was spread on glass slides and solidified at 4 °C for 10 min. Cells were harvested 48 h after RITA treatment, mixed with low-melting-point agarose (LMA), and plated on glass slides with NMA as the second layer. The glass slides with agarose gel were placed in pre-cooled lysis buffer for 2 h at 4 °C, subjected to electrophoresis at 25 V for 30 min under alkaline conditions, and stained with PI. The presence of comet tails was determined using U-HGLGPS (Olympus, Japan). The tail moment was calculated as (percentage of DNA in the tail) × (tail length), where the percentage of DNA in the tail and tail length were quantified using the CASP software.
8
ARID1A Knockdown Using siRNA
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Two different ARID1A siRNAs (siARID1A #1 and siARID1A #2) were designed and synthesized by RiboBio (Guangzhou, China). The sequences were as follows: siARID1A #1,5′-GGACCTCTATCGCCTCTAT-3′; siARID1A #2,5′-GAAGCAGGCACCACTAACT-3′. siRNA transfection was performed using Lipofectamine 3000, and the procedure strictly followed the manufacturer’s instructions. Reverse transfection was performed to silence ARID1A in a 12-well plate. Briefly, after cells were trypsinized, 5 μL of siRNA was pipetted into 100 μL Opti medium to dilute to the indicated concentration and then mixed with 100 μL Opti medium containing 2 μL Lipofectamine 3000, and the transfection mixture was incubated at room temperature for 20 min. Cells were trypsinized and 1000 μL of cell suspension (1 × 105 cells per well) was added to each well containing the transfection mixture. The cells were then incubated for 72 h in a CO2 incubator, and cell images were captured using U-HGLGPS (Olympus, Japan). ImageJ software was used to measure cell density and assess cell viability. The silencing efficiency was assessed by immunoblotting using an anti-ARID1A antibody.
9
Quantifying Intracellular ROS in MCF-7 Cells
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The intracellular ROS levels in the MCF-7 cells were measured using 2′,7′-dichlorofluorescein diacetate (DCFH-DA). The DCFH-DA probe is taken up by cells, deacetylated to generate 2′, 7′-dichlorodihydrofluorescein (DCFH), and then rapidly oxidized to generate a strong fluorescent product 2′,7-dichlorofluorescein (DCF). Briefly, MCF-7 cells were cultured overnight in 96-well plates in 5% CO2 at 37 °C. After treatment, the cells were incubated with 10 µM DCFH-DA for 30 min at 37 °C. After incubation, the cells were rinsed with serum-free MEM. Fluorescent images of the cells were acquired using a fluorescence microscope (Olympus U-HGLGPS, Suzhou, China).
10
Edu-Based DNA Synthesis Assay
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Edu (5-ethynyl-2′-deoxyuridine), a novel thymidine analogue, can be incorporated into newly synthesized DNA instead of thymidine during DNA synthesis. After treating the MCF-7 cells with various concentrations of extract, they were thoroughly rinsed thrice using PBS. Next, the cells were exposed to a working solution of Edu (10 µM) and incubated in darkness for 4 h at a temperature of 37 °C. After incubation, the cells were washed with PBS three times and fixed with 4% paraformaldehyde for 15 min. Next, the cells were permeabilized with 0.3% TritonX-100 for 15 min and washed with PBS three times. The cells were then incubated with Hoechst 33342 for 10 min. The images were captured using a fluorescence microscope (Olympus U-HGLGPS, Shanghai, China).
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