Cell motility was assessed in Boyden chambers as described [80 (link),81 (link),109 (link)]. Briefly, HTS FluoroBloks™ inserts (Becton Dickinson) were saturated with PBS-1% bovine serum albumin for 2 h at room temperature. Serum-starved cells were labeled with DiI (Molecular Probes) for 20 min at 37 °C and then seeded in the HTS FluoroBloks™ upper chamber in either SFM or SFM supplemented with either 1% FBS, 10 nM of IGF-I, IGF-II or insulin and incubated at 37 °C for either 4 (1% serum) or 18 h. After fixing in 4% paraformaldehyde, membranes were mounted on a slide, and migrated cells were counted and photographed with a Zeiss Axiovert 200M live-cell microscope at the Kimmel Cancer Center Confocal Microscopy Core Facility. Anchorage-independent grow was measured by colony formation in soft-agar as previously described [80 (link),81 (link),110 (link),111 (link)]. Briefly, cells were seeded in soft-agar at a density of 5 × 103 cells/35 mm plate and counted after three weeks in culture. Colonies > 150 μm were scored as positive.
Hts fluorobloks inserts
Manufactured by BD
Sourced in United States
HTS FluoroBloks™ inserts are a type of laboratory equipment used for high-throughput screening applications. They are designed to provide a controlled and consistent environment for fluorescence-based assays. The inserts feature a black, opaque bottom that minimizes background fluorescence, allowing for improved signal-to-noise ratios in fluorescent experiments.
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Lab products found in correlation
3 protocols using hts fluorobloks inserts
1
Assessing Cell Motility and Anchorage-Independent Growth
2
Cell Migration and Invasion Assay
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HTS FluoroBloks™ inserts (Becton Dickinson, Durham, NC, USA) were saturated with PBS-1% bovine serum albumin for 2 h at room temperature. Serum-starved cells were labeled with DiI (Molecular Probes, Grand Island, NY, USA) for 20 min at 37°C and seeded in the HTS FluoroBloks™ upper chamber in SFM or SFM supplemented with progranulin (40 nM) and incubated at 37°C for 18 h. Membranes were fixed in 4% paraformaldehyde, mounted on slides and migrated cells were counted and photographed with a Zeiss Axiovert 200 M cell live microscope. Cell invasion was assessed by using BD Matrigel™ Invasion Chambers (BD Biocoat, Bedford, MA, USA) with 8.0 μm filter membranes. Cells (5 × 104) in 200 μl of SFM were plated onto each filter, and 750 μl of SFM or SFM supplemented with progranulin (40 nM) in the lower chamber. After 24 h filters were washed, fixed, and stained with Coomassie Brilliant Blue. Cells on the upper surface of the filters were removed with cotton swabs. Cells that had invaded to the lower surface of the filter were counted under the microscope.
3
Cell Motility and Anchorage-Independent Growth Assays
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