Cell invasion was evaluated by a modified Boyden chamber method using a 48-well chemotaxis chamber (NeuroProbe, Gaithersburg, MD). Briefly, 25,000 HepG2 and 20,000 Hep3B cells per well were seeded onto an 8 µM pore PDVF membrane (NeuroProbe) previously treated with collagen type IV (BD Biosciences, San Jose, CA) and conditioned medium from scramble- and siPRPF8-treated cells. The lower well was filled with FBS-free medium (negative control) or medium containing 10% FBS, with two replicates per condition23 (link). After 24 h, the non-migrated cells were removed, and the migrated cells were fixed with crystal violet solution (6% glutaraldehyde, 0.5% crystal violet). Images of random fields were acquired and analyzed with ImageJ. The experiments were performed in triplicate.
48 well chemotaxis chamber
Manufactured by Neuro Probe
Sourced in United States
The 48-well chemotaxis chambers are a laboratory equipment designed to study cell migration and chemotaxis, a process where cells move in response to chemical gradients. The chambers provide a controlled environment for observing and quantifying cell movement. Each chamber contains 48 individual wells, allowing for multiple experimental conditions to be tested simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
Collagen type 4, by BD (1 mentions)
Giemsa solution, by Bio-Optica (1 mentions)
Axioskop 20, by Zeiss (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Easysep mouse monocyte enrichment kit, by STEMCELL (1 mentions)
Human il 8, by Merck Group (1 mentions)
Polycarbonate filter, by Neuro Probe (1 mentions)
Type 1 collagen, by BD (1 mentions)
Osmonics labstore, by GE Healthcare (1 mentions)
F1141, by Merck Group (1 mentions)
Pherastar plate reader, by BMG Labtech (1 mentions)
17 protocols using 48 well chemotaxis chamber
1
Cell Invasion Assay with Boyden Chamber
2
Chemotaxis Assay for hfNBM Migration
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Cell migration was assessed using a 48-well chemotaxis chamber (Neuro Probe, Cabin John, MD, USA) with polycarbonate membranes (pore size 8 µm; Neuro Probe; #PFB8) coated with collagen type I (20 µg/mL, Sigma-Aldrich Corp.; #CC050). Briefly, the lower wells of the chamber were filled with serum-free or 10% FBS culture medium to evaluate basal or induced hfNBM migration, respectively. TNFα pre-treated (10 ng/mL for 48 h) or untreated hfNBMs (4 × 104 cells) were seeded in serum starved condition into the upper well and incubated for 6 h at 37 °C in 5% CO2 atmosphere. After the incubation period, the migrated cells in the lower side of the membrane were fixed with absolute methanol for 15 min, washed with PBS, and stained for 30 min with 10% Giemsa solution (BioOptica, Milan, Italy; #05-12005E) in PBS. No migrated cells, in the upper part of the membrane, were removed and the filter was mounted on a glass slide for visualization. The number of migrated hfNBMs was counted in blind under an optical microscope (Zeiss Axioskop 20; Carl Zeiss S.p.A., Milan, Italy) in 10 fields for each well. Each experimental point was replicated at least six times in three independent experiments.
3
Chemotaxis Assay for Colorectal Cancer
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Chemotaxis assays were performed using a 48-well chemotaxis chamber (Neuro Probe, Gaithersburg, MD) according to the published procedures32 (link). For SW480 and HT29 cells, after 12 h incubation with fMLF (1 μM), with or without a 15 min pretreatment with cyclosporine H (1 μM) or Boc1 (10 μM) at 37 °C, the filter was removed and fixed with methanol and stained with 0.1% crystal violet. Cells were counted in three random high-power fields in triplicate samples. The results were expressed as chemotaxis index which represents the fold increase in the number of migrated cells in response to chemoattractants over the response to control medium.
4
Yacon Extract's Effect on Glioma Cell Migration
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Boyden chamber assay was performed as previously reported [15 (link)]. To determine the effect of yacon extract on the migration of C6 glioma cells, the Boyden chamber assay was performed in a 48-well chemotaxis chamber (Neuro-Probe, Gaithersburg, MD, USA). Briefly, polycarbonate filter membranes (Neuro-Probe) with a pore size of 8 µm were coated with 0.1% collagen type-I and dried at room temperature for 1 h. Two different concentrations of yacon extract (200 and 300 µg/mL in 10% FBS media) were prepared and then 27.8-µL portions were dispensed into the lower chamber of the wells. The lower chamber containing the extract was then covered by the coated membrane, and 50 µL of C6 glioma cells (in serum-free media) were loaded into the upper chamber at a concentration of 1 × 106 cells/mL. Following culture at 37℃ in 5% CO2, cells on the lower surface of the membrane were fixed with methanol and stained using Diff-Quick. After staining, cells that had migrated through the filter membrane were imaged and counted using an inverted microscope as described above.
5
Neutrophil Chemotaxis Assay Protocol
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Whole blood was collected in heparin tubes. Neutrophils were enriched by gradient centrifugation over Ficoll-Paqueplus (GE Healthcare) within 6 h of blood collection. Red blood cells were lysed with ACK (ammonium-chloride-potassium) lysis buffer, and the remaining neutrophils were washed with Hanks balanced salt solution (HBSS) supplemented with 0.05% heat-inactivated fetal bovine serum (FBS), and resuspended at a concentration of 107 cells/ml in RPMI 1640 supplemented with 10% FBS.
Neutrophil chemotaxis was assessed using a 48-well chemotaxis chamber (NeuroProbe). RPMI 1640, fMLP (100 nM), or IL-8 (100 ng/ml) was loaded into the bottom of the chamber, a polycarbonate filter with 5-µm pores was laid down, and 105 neutrophils in 50 µl of RPMI 1640 was loaded into the top of the chamber. After 1 h of incubation at 37°C, the filter was removed, fixed, and stained. Densitometry analysis was performed to determine the relative amount of neutrophil chemotaxis across the membrane toward the bottom chamber.
Neutrophil chemotaxis was assessed using a 48-well chemotaxis chamber (NeuroProbe). RPMI 1640, fMLP (100 nM), or IL-8 (100 ng/ml) was loaded into the bottom of the chamber, a polycarbonate filter with 5-µm pores was laid down, and 105 neutrophils in 50 µl of RPMI 1640 was loaded into the top of the chamber. After 1 h of incubation at 37°C, the filter was removed, fixed, and stained. Densitometry analysis was performed to determine the relative amount of neutrophil chemotaxis across the membrane toward the bottom chamber.
6
Inhibition of Glioma Cell Migration
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Boyden chamber assay was performed using a 48-well chemotaxis chamber (Neuro-Probe, Gaithersburg, MD, USA) to confirm the inhibitory effect of CK on migration of C6 glioma cells. Scratch wound healing analyses were performed as previously reported [25 (link)]. Briefly, C6 glioma cells were suspended in serum-free medium at a density of 1 × 106 cells/ml, and 50 µl of the cell suspension was loaded in the upper chamber wells. Treatment with SDF-1 with or without CK at different concentrations was performed in the lower chamber with incubation at 37℃ and 5% CO2 atmosphere for 90 min. Cells on the lower surface of the membrane were fixed and stained using Diff Quick solutions. After staining, the migrated cells were photographed and counted under a microscope (Nikon, Tokyo, Japan). The migrated cells were measured using Image J software.
7
Monocyte Migration Assay Using fMLP
8
Neutrophil Chemotaxis Assay
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Chemotaxis assay was performed as described by Moyes et al. (2009) with minor modifications. In triplicate, 30 μL of either RPMI medium (nonrandom migration) or human IL-8 (Sigma Aldrich Co., 100 ng/ mL, specific migration) was added to the bottom wells of a 48-well chemotaxis chamber (Neuro Probe, Gaith-ersburg, MD). Fifty microliters of cell suspension (3 × 10 6 PMNL/mL) was added to the top of a preloaded filter (Fisherbrand, Thermo Fisher Scientific Inc.) . Plates were incubated for 60 min at 37°C, 5.0% CO 2 , and 95% relative humidity. Five fields per well were counted using a 100× magnification to determine the number of PMNL that migrated in response to IL-8 or RPMI. An average of the 5 fields per well was calculated. For statistical analysis, a chemotaxis index was calculated by subtracting the random migration (RPMI) from the migration with IL-8.
9
T Cell Migration Quantification
10
T Cell Migration Quantification
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T cell migration was measured using 48-well chemotaxis chambers (Neuro Probe, Inc.) equipped with 5 μm-pore polycarbonate membranes which were coated with fibronectin (1 μg/ml). Cells (1 × 106/ml) were added to the upper chamber. Migration was assessed after 2 h for TILs and CD8+ T cells at 37°C in humidified air containing 5% CO2. Migrated cells were fixed in Diff-Quik Fix and stained using Diff-Quik II. Migration was quantified by color intensity measurements using ImageJ software.
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