Migration assay was performed as previously described (Nangia-Makker, Vitaly, & Avraham, 2012 (link); do Prado et al., 2017 (link)). Briefly, bovine adrenal medullary endothelial cells (BAMEC) maintained in EMEM containing 10% FBS were pre-labeled with DiL (green) and incubated in one well of a 2-well culture-insert chamber (2.4 × 104 cells/well). HCT116, HT29 or PC3 cells prelabeled with DiO (red) were incubated in the other wells of the culture-insert chamber (2.4 × 104 cells/well). After 12 h, the cells were washed with PBS and the culture-insert chamber was removed. Cells were treated or not with MCP, MCP fractions or lactose for 24 h. Migration of co-cultures toward each other was observed after 24 h using a LSM 510 META LNO Laser Scanning Microscope (Carl Zeiss, Oberkochen, Germany; The Wayne State University Microscopy and Imaging Core Facility) and migration was compared to the co-culture before treatment (0 h).
Lsm 510 meta lno laser scanning microscope
Manufactured by Zeiss
Sourced in Germany
The LSM 510 META LNO Laser Scanning Microscope is a high-performance optical microscope system designed for advanced imaging applications. It utilizes a laser light source and scanning mirrors to capture detailed images of samples. The system is capable of providing high-resolution, three-dimensional imaging capabilities.
Automatically generated - may contain errors
2 protocols using lsm 510 meta lno laser scanning microscope
1
Co-culture Migration Assay for Endothelial and Cancer Cells
2
Immunofluorescence Analysis of hBD4 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect hBD4 expression, MDBK cells (1.0 × 105 cells per well) were cultured directly on the 24-well plates with glass coverslips and infected with the rNDV-hBD4 at an MOI of 1 for 1 h. At 12 hpi, Golgi stop protein transport inhibitor (BD Bioscience, San Jose, CA) was treated for 10 h. The cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100 in PBS (PBST). After treated with PBST containing bovine serum albumin (PBST-BSA), NDV proteins and expressed hBD4 were detected primarily with polyclonal rabbit anti-NDV antibody and monoclonal mouse anti-hBD4 antibody, respectively. Then, Texas Red conjugated anti-rabbit antibody (Jackson ImmunoResearch, West Grove, PA) and Alexa 488 conjugated anti-mouse antibody (Invitrogen) were used as secondary antibodies. Hoechst 33258 (Invitrogen) was used for cell nucleus detection. The cells were imaged with a LSM 510 Meta LNO laser scanning microscope (Carl Zeiss, Jena, Germany) at Korea Basic Science Institute, Chuncheon, Korea. PBS-infected samples were used as a control.
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!