To determine the effects of GRP78 KD on PC3 cells adhesion to bone, PC3 cells were transfected with GRP78 siRNA and harvested 48 h later and cultured with hFOB 1.19 cells plated in a 96 -well plate until confluent. To track the PC3 cells in co-culture, transfected cells were labeled with the Vybrant CFDA SE (green) Cell Tracer Kit (5 μM) (Thermo Fisher Scientific) counted on a Z2 particle counter and size analyzer (Beckman Coulter), and then seeded at 10,000 cells/well onto confluent OSB. For comparison, parallel co-cultures were treated overnight with 20μg/mL of a N-cad NAb, clone CG-4 (Sigma-Aldrich). Number of fluorescently labeled cells [Ncoculture] were counted by high content screening (Cell Insight CX5, Thermo Fisher Scientific) following an 18 h co-incubation period. Supernatants were then transferred from co-culture wells into empty ones to determine the number of fluorescently labeled floating cells [Nsupernatant]. Adherence was calculated as %Nsupernatant/ Nco-culture.
Z2 particle counter and size analyzer
Manufactured by Beckman Coulter
Sourced in United States
The Z2 particle counter and size analyzer is a laboratory instrument designed to measure the size and count of particles in a sample. It utilizes the Coulter Principle to provide accurate and reproducible particle analysis data.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using z2 particle counter and size analyzer
1
Investigating GRP78 KD Effects on PC3-Bone Adhesion
2
Cell Volume Measurement Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell volumes were determined using a Beckman Z2 particle counter and size analyzer with filtering criteria set to include cells between 10 and 30 μm.
3
Bronchoalveolar Lavage Fluid Collection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Trachea was exposed through midline incision and intubated with aseptic 22 Abbocath-T catheter (Abbott, Sligo, Ireland). BAL was performed by injecting two parts of 0.5 mL aseptic saline into the right lung. The recovered BALF (0.8 mL) was rotated at 4 °C for 10 min at 260×g, and the precipitate was re-suspended in 0.5 mL sterile PBS. The total number of cells was counted by a Z2 Particle Counterand Size Analyzer (Beckman-Coulter, Miami, FL, USA). Differential cell counting (Diff-Quick, Baxter, UK) was performed on cell centrifuge smears stained with modified Giemsa stain.
4
Cell Morphology Analysis via Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cell morphology analysis, PC3 cells were transfected with GRP78 siRNA and harvested 48 h later or treated overnight with 20μg/mL of a N-cad neutralizing monoclonal antibody, clone CG-4 (N-cad NAb, Sigma-Aldrich) prior to analysis. The cells were counted on Z2 Particle Counter and Size analyzer (Beckman Coulter) and re-seeded at 20,000 cells/well in a standard 96 well plate and left to culture for an additional 18 h. The wells were subsequently washed and imaged under bright field on a Cell Insight CX5 high content screening instrument (Thermo Fisher Scientific). Images were analyzed using ImageJ software package (NIH). 15 fields of view (5 each from 3 independent experiments) of control cells or cells treated with GRP78 siRNA were analyzed using the ImageJ software package (NIH). Cells with a clearly defined spherical and darker border (under bright field) were considered as rounded. Morphology was calculated as rounded: Nrounded cells/Ntotal cells or elongated: [Ntotal cells-Nrounded cells]/Ntotal cells and represented as a mean percentage ± SD.
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!