How can I increase the transduction efficiency with the BacMam 2.0 reagents such as the the CellLight and Premo products?
Try varying particle-to-cell ratio (PPC), incubation volume, temperature and, cell density (if adherent cells are transduced). For adherent cells, we recommend a confluence of about 70%. Following the PPC, adjusting the volume is the next best parameter to change to optimize protein expression. If that doesn't work, you can also use the BacMam Enhancer Kit (Cat. No. B10107).
Substrate stiffness regulates cadherin-dependent collective migration through myosin-II contractility.
Authors:Ng MR, Besser A, Danuser G, Brugge JS,
Journal:J Cell Biol
PubMed ID:23091067
'The mechanical microenvironment is known to influence single-cell migration; however, the extent to which mechanical cues affect collective migration of adherent cells is not well understood. We measured the effects of varying substrate compliance on individual cell migratory properties in an epithelial wound-healing assay. Increasing substrate stiffness increased collective cell ... More
Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines.
Authors:Vercauteren D, Deschout H, Remaut K, Engbersen JF, Jones AT, Demeester J, De Smedt SC, Braeckmans K,
Journal:ACS Nano
PubMed ID:21923168
'To gain a better understanding of intracellular processing of nanomedicines, we employed quantitative live-cell fluorescence colocalization microscopy to study endosomal trafficking of polyplexes in retinal pigment epithelium cells. A new, dynamic colocalization algorithm was developed, based on particle tracking and trajectory correlation, allowing for spatiotemporal characterization of internalized polyplexes in ... More
Intracellular trafficking mechanism, from intracellular uptake to extracellular efflux, for phospholipid/cholesterol liposomes.
Authors:Un K, Sakai-Kato K, Oshima Y, Kawanishi T, Okuda H,
Journal:Biomaterials
PubMed ID:22858002
'Liposomes are widely used as drug delivery vehicles to transfer chemotherapeutic agents, proteins, and nucleic acids into target cells. To improve therapeutic effects and reduce unexpected toxic side-effects, it is necessary to understand the mechanism of liposomal uptake into cells, and the intracellular fate of internalized liposomes. The intracellular fate ... More
Glycophthalocyanines as photosensitizers for triggering mitotic catastrophe and apoptosis in cancer cells.
Authors:Soares AR, Neves MG, Tomé AC, Iglesias-de la Cruz MC, Zamarrón A, Carrasco E, González S, Cavaleiro JA, Torres T, Guldi DM, Juarranz A,
Journal:Chem Res Toxicol
PubMed ID:22394248
'Photodynamic therapy (PDT) is a treatment modality for different forms of cancer based on the combination of light, molecular oxygen, and a photosensitizer (PS) compound. When activated by light, the PS generates reactive oxygen species leading to tumor destruction. Phthalocyanines are compounds that have already shown to be efficient PSs ... More
A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and Structures, Part III: Reagents for Actin, Tubulin, Cellular Membranes, and Whole Cell and Cytoplasm.
Authors:Kilgore JA, Dolman NJ, Davidson MW,
Journal:
PubMed ID:24510770
'Non-antibody commercial fluorescent reagents for imaging of cytoskeletal structures have been limited primarily to tubulin and actin, with the main factor in choice based mainly on whether cells are live or fixed and permeabilized. A wider range of options exist for cell membrane dyes, and the choice of reagent primarily ... More