The mucosal layer separated from the excised mouse bladder was washed with PBS and then settled in the Ussing chamber device with a constant mucosal area of 0.1 cm2. The urothelial mucosa was fixed to the feeding tank of the chamber. The feeding tank was filled with Tyrode's solution (Solarbio, Beijing, China) containing Cy5.5-labeled PLL/pEGFP or PLLF/pEGFP, whereas the reception tank was filled with fresh Tyrode's solution. The Ussing chamber was maintained at 37 ℃ and supplied with O2/CO2 (95/5%). 100 μL Tyrode solution in the reception tank was pipetted out after 15, 30, and 60 min for Cy5.5 signal detection using a Spark® Multimode Microplate Reader (TECAN, Switzerland). The resulting value for the PLL/pEGFP group after 15 min was used as the control to calculate the relative penetration efficiency.
Tyrode s solution
Manufactured by Solarbio
Sourced in China
Tyrode's solution is a balanced salt solution commonly used in biological and medical research. It is a physiological saline solution that mimics the ionic composition of extracellular fluid, maintaining the appropriate osmotic pressure and pH for cell and tissue experiments.
Automatically generated - may contain errors
Lab products found in correlation
Soybean phospholipid, by Yuanye Bio-Technology (1 mentions)
Fecl3 6h2o, by Maclin Biochemical Technology (1 mentions)
Methylene blue, by Merck Group (1 mentions)
Bca kit, by Beyotime (1 mentions)
Red blood cell lysis buffer, by Solarbio (1 mentions)
Cell counting kit 8 cck 8, by Targetmol (1 mentions)
Fitc dextran, by Xi’an Ruixi (1 mentions)
Tcs5 sp5, by Leica (1 mentions)
Pluronic f 127, by Thermo Fisher Scientific (1 mentions)
Fluo 4 am, by Thermo Fisher Scientific (1 mentions)
Fluo 4 am, by Beyotime (1 mentions)
Spark multimode microplate reader, by Tecan (1 mentions)
Giemsa, by Beyotime (1 mentions)
7 protocols using tyrode s solution
1
Urothelial Permeability Evaluation
2
hiPSC-CMs Characterization and Cardioprotection
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hiPSC-CMs were obtained from Help Therapeutics (Nanjing, China). Danhong injection was purchased from Buchang Pharma (Shandong, China), and aconitine and sotalol were purchased from Desite (Chengdu, China) and Merck (Darmstadt, Germany), respectively. Tyrode's solution was obtained from Solarbio (Beijing, China). There are no medical ethical issues involved in this study.
3
Synthesis and Characterization of Nanoparticles
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FeCl3·6H2O were purchased from Macklin; 3,4,5-Trihydroxybenzoic acid was purchased from Shanghai D&B Biological Science and Technology Co., Ltd; ACD anticoagulant was purchased from Shanghai Zhuocai Technology Co., Ltd; Tyrode's solution and red blood cell lysis buffer were purchased from Solarbio; Phenylmethylsulfonyl fluoride (PMSF), polyvinyl pyrrolidone (PVP, MW 8000), and cholesterol were purchased from Aladdin; soybean phospholipid was purchased from Shanghai Yuanye Bio-Technology Co., Ltd; 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-poly(2-ethyl-2-oxazoline) (DSPE-PEOz, MW 2000), 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol (DSPE-PEG, MW 2000) and FITC-Dextran were purchased from Xi'an Ruixi Biological Technology Co., ltd; PKH67 and PKH26 fluorescent dye were purchased from Beijing Bio Rab Technology Co. Ltd; IR780 was purchased from Alfa Aesa; Methylene blue was purchased from sigma; H2O2 was purchased from Sinopharm Chemical Reagent Co., Ltd; BCA kit was purchased from Beyotime Biotechnology; and Cell Counting Kit-8 (CCK-8) was purchased from TargetMol, USA.
4
Colon Smooth Muscle Contractility Assay
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Rats were harvested at moribund. Colon tissues were dissected free, cut transversely into 2 cm muscle strips. Each strip was then incubated in the HW200S smooth muscle homeothermic system (Tai Meng Technology Co., Ltd., China) containing 37°C Tyrode's solution (Solarbio, China). All solutions were continuously aerated by a mixed gas (95% O2/5% CO2). The upper and lower ends of each strip were threaded diagonally and fixed on a tension transducer (FT-102 N, Tai Meng Technology Co., Ltd., China) and the L-shaped hook, respectively. Contractile activities were recorded by a BL-420N laboratory system (Tai Meng Technology Co., Ltd., China) by connecting with the tension transducer.
5
Intracellular Calcium Dynamics in hESC-CMs
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hESC‐CMs were singularized and seeded into a confocal dish or chamber (Sigma). Fluo‐4 AM (Beyotime) mother liquor was diluted to 5 µmol/L working solution with PBS for next cell loading. After washing three times with PBS, the cells were incubated with Fluo‐4 AM working solution at 37°C for 15 minutes during which 0.02% Pluronic F‐127 (Invitrogen) can be added to promotes Fluo‐4 AM into cardiomyocytes. Then, the cells were washed three times with PBS, and the Fluo‐4 fluorescence intensity was measured by a laser confocal microscope (Leica, TCS5 SP5, Germany) to determine the intracellular Ca2+ changes. The total scan time is 32.768 seconds, collecting four pictures, each for 8.192 seconds. Subsequently, a caffeine release experiment was performed using calcium‐free Tyrodes solution (Solarbio, China) to deplete Ca2+ in the extracellular fluid and 10 mmol/L caffeine to induce sarcoplasmic reticulum Ca2+ release. Data analysis was performed by ImageJ and Igor software.
6
Cauda Epididymal Sperm Quality Evaluation
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Cauda epididymis was dissected from WT, SKO, and DKO male mice (10 weeks old, n = 5) and transferred to Tyrode’s solution (T1421-500 ml: Solarbio, Beijing, China). After incubating the samples at 37 °C for 10 min, sperm quality was measured using the CASA system of the Hamilton Thorne CEROS II (Medealab™, Erlangen, Germany).
7
Sperm Extraction and Analysis from Mouse
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The cauda epididymis was dissected from an individual mouse, and then incubated in Tyrode’s Solution (Solarbio) at 37 °C for 15 min to release the sperms. The supernatant was collected, and sperm counts and motility were evaluated using the Computer Assisted Semen Analysis (CASA) system (Hamilton). Otherwise, the supernatant was centrifuged at 1000 g for 15 min, the sperm pellet was resuspended in 4% paraformaldehyde, and spread on precoated slides. For morphological observation, Giemsa staining was performed according to the manufacturer's instructions (Beyotime). At least 200 sperms were recorded for each sample.
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