Lab-Tek II 4-well chamber slides (Nalge Nunc International, Naperville, IL) were coated with 100 μl of reduced growth factor basement membrane extract (BME; Trevigen, Gaithersburg, MD) which solidified for 30 minutes at 37°C. Cells were then seeded onto the solidified BME layer at a density of 5,000 cells/well in medium containing 10% FBS and 2.5% BME. Medium was refreshed every three days. Cell colony sizes were determined with ImageJ software from a total of 4 images from each of duplicate wells for each experiment.
Lab tek 2 4 well chamber slides
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The Lab-Tek II 4-well chamber slides are a type of lab equipment designed for cell culture and microscopy applications. They provide a convenient and controlled environment for growing and observing cells in vitro. The slides feature four separate chambers, each with its own glass surface, allowing for the simultaneous culture and examination of multiple samples.
Automatically generated - may contain errors
Lab products found in correlation
Bcecf am, by Thermo Fisher Scientific (2 mentions)
Lsm 710 laser scanning microscope, by Zeiss (2 mentions)
Lsm 780 laser scanning confocal microscope, by Zeiss (1 mentions)
Pflag cmv2 vector, by Merck Group (1 mentions)
Fluorescein isothiocyanate (fitc), by Dojindo Laboratories (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Fv10i, by Olympus (1 mentions)
Endothelial cell basal medium mv, by PromoCell (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Hcaec, by PromoCell (1 mentions)
Smooth muscle cell basal medium, by Lonza (1 mentions)
6 protocols using lab tek 2 4 well chamber slides
1
3D Cell Culture on Basement Membrane
2
Quantifying Apoptotic Blebbing Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
THP-1 and Jurkat cells were stained with MitoTracker Green (100 nM) and Hoechst 33342 (5 μg/ml) before induction of apoptosis. Jurkat and HL-60 cells were stained with Hoechst 33342 (5 μg/ml) and SYTO RNAselect green (500 nM). Samples were incubated in Lab-TekII 4-well chamber slides (Nunc) and confocal microscopy was performed at 37 °C in a humidified atmosphere with 5% CO2 using Zeiss LSM780 Laser Scanning Confocal Microscope (Carl Zeiss SAS, Jena, Germany) to monitor the progression of ApoBD formation within 4 hours.
3
Visualizing AIM Binding to CD36
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pFLAG-CMV2 vector (Sigma-Aldrich) was used to generate expression vectors for mouse CD36. Mouse and feline rAIM were labelled with FITC (Dojindo) and used for experiments. HEK293T cells were cultured in Lab-Tek II 4-well chamber slides (Nunc) 2 days before the assay, and transfected with mouse CD36 expression vector 1 day before the assay. rAIM was added to each well at 20 µg/mL and incubated for 30 min at 37 °C. Then, the cells were fixed in 4% paraformaldehyde (PFA) in PBS for 15 min at room temperature and stained for CD36. The nuclei were stained with Hoechst 33342 and mounted in Prolong Gold Antifade reagent (Thermo Fisher Scientific). Incorporation of AIM was analysed by confocal microscopy (FV10i; Olympus, Tokyo, Japan).
4
Culturing Coronary Artery Endothelial and Smooth Muscle Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human coronary artery endothelial cells (HCAEC, PromoCell, Heidelberg, Germany) were cultured in Endothelial cell basal medium MV (PromoCell) supplemented with the Supplement Pack Endothelial Cell GM MV (PromoCell) comprising fetal calf serum, endothelial cell growth supplement, recombinant human epidermal growth factor, heparin, and hydrocortisone; as well as 100 U/ml penicillin and 100 μg/mL streptomycin (Lonza, Basel, Switzerland) under standard cell culture conditions (37 °C, 5% CO2). For microscopy, 6 × 104 cells were seeded into each well of Lab-Tek II 4-well Chamber slides (Nunc A/S, Roskilde, Denmark), cultured until they reached 80–95% confluency, and fixed with 4% PFA for 15 min and kept in PBS until the time of staining.
Human coronary artery smooth muscle cells (CaSMC, Clonetics, Lonza) were cultured under standard cell culture conditions (37 °C, 5% CO2) in Smooth Muscle cell Basal Medium (Clonetics Lonza) supplemented with SmGm-2 SingleQuots supplement pack containing epidermal growth factor, fibroblast growth factor, insulin, gentamicin sulfate, and fetal bovine serum; as well as 100 U/ml penicillin and 100 μg/ml streptomycin (Lonza). For microscopy, 2.5 × 104 cells were seeded into each well of Lab-Tek II 4-well Chamber slides (Nunc), cultured until they reached 80–95% confluency, and fixed with 4% PFA for 15 min and kept in PBS until the time of staining.
Human coronary artery smooth muscle cells (CaSMC, Clonetics, Lonza) were cultured under standard cell culture conditions (37 °C, 5% CO2) in Smooth Muscle cell Basal Medium (Clonetics Lonza) supplemented with SmGm-2 SingleQuots supplement pack containing epidermal growth factor, fibroblast growth factor, insulin, gentamicin sulfate, and fetal bovine serum; as well as 100 U/ml penicillin and 100 μg/ml streptomycin (Lonza). For microscopy, 2.5 × 104 cells were seeded into each well of Lab-Tek II 4-well Chamber slides (Nunc), cultured until they reached 80–95% confluency, and fixed with 4% PFA for 15 min and kept in PBS until the time of staining.
5
CSR1 Fluorescence Imaging in pH Sensitivity
6
CSR1 Fluorescence Imaging in pH Sensitivity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were grown in Lab-Tek II 4-well chamber slides (Thermo Scientific) and 2 wells were stained with 2 μM of CSR1 in DMEM for 10 min. Cells were then washed twice with 500 μL Buffer A (20 mM MOPS, 140 mM NaCl, 150 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, pH 7.4 at 37 °C) and then allowed to equilibrate for 30 min at 37 °C in Buffer A containing 10 μM nigericin and 10 μM valinomycin. Confocal microscopy images (4 positions per well) were acquired on a Zeiss LSM 710 laser scanning microscope with a 20X objective lens. CSR1 was excited at 594 nm with a He-Ne laser, and emission was collected using a META detector between 610 and 700 nm. The buffer in one well was changed to Buffer A (pH 7.3) containing nigericin and valinomycin, whereas the buffer in the second well was changed to Buffer A containing nigericin and valinomycin, but at pH 6.7. After equilibration at 37 °C for ~30 min, the same positions were imaged again. Control experiments using the pH indicator 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester (BCECF-AM, Life Technologies) showed that this was sufficient time for intracellular pH to stabilize. Images were analyzed using ImageJ (National Institutes of Health), with the initial and final fluorescence for each position compared. The experiment was carried out twice.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!