Scintillant Bioscience

Would you like to learn more about our contract-research services for drug discovery and development? You can book a discovery call to learn more at the following link: https://lnkd.in/gdGx_AKj

Ask us about our contract-research services for high-content assay development, screening and analysis. Please click on the link to book a meeting to learn more: https://lnkd.in/gdGx_AKj

Let us help you with your therapeutics discovery and development challenges. Please download this short slide deck that summarizes our capabilities and then book a call with us at the following link if you would like to learn more. https://lnkd.in/gdGx_AKj

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Please download this brochure, which summarizes Scintillant Bioscience's capabilities and scientific services. To learn more, please book a free discovery call with this link: https://lnkd.in/gdGx_AKj

If you would like to learn more about our contract-research services at BIO 2024, you can schedule a meeting with our CEO, Russ Teichert, through the BIO One-on-One Partnering system. He will be participating in One-on-One Partnering at the BioUtah booth (Utah Pavilion, # 4017).

Tips and Tricks for Cell-Based Imaging Assays: Immunocytochemistry Permeabilization-Mild Detergents Saponin and digitonin are milder membrane solubilizers compared to Triton X-100 and NP-40. They can be used at concentrations of 0.1% to 0.5% for up to 30 minutes. These detergents create pores of sufficient size for antibody penetration without dissolving the plasma membrane. Saponin and digitonin bind to cholesterol in the plasma membrane, forming pores that serve as gateways for antibody entry. This approach is particularly advantageous when preserving delicate cytoplasmic structures and maintaining the integrity of intracellular membranes is critical for accurate analysis. However, it is worth noting that these detergents may be less effective in permeabilizing certain cell types compared to other permeabilization agents. One significant advantage of saponin and digitonin is their selective permeabilization of the plasma membrane while leaving intracellular membranes intact. It is important to emphasize that saponin and digitonin are not recommended for permeabilizing the nuclear membrane. Tween-20 is a mild detergent that is occasionally used for permeabilizing fixed cells but is more commonly employed in blocking and binding buffers to help prevent non-specific antibody binding. #CellImaging #Immunocytochemistry #Permeabilization #CellBiology #Saponin #Digitonin #TritonX100 #NP40 #LabTips #CellAssays #Biotech #ResearchTips #ScienceTips #LabTechniques #Microscopy #CellCulture

Tips and Tricks for Cell-Based Imaging Assays: Immunocytochemistry Permeabilization-Strong Detergents Following fixation, the crucial next step in the ICC workflow is cell permeabilization, which allows antibodies to access their target antigens within the cell. Various approaches to cell permeabilization exist, each with its advantages and limitations. For example, Triton X-100 and NP-40 are non-ionic detergents commonly employed for cell permeabilization. They effectively disrupt the plasma membrane by solubilizing lipids and creating transient pores. These detergents are efficient in permeabilizing most cell types, enabling antibodies to penetrate the cell and bind to target antigens. They also partially dissolve the nuclear membrane, making them suitable for nuclear antigen staining. Typically, they are used at concentrations ranging from 0.1% to 0.2%, and in some cases up to 0.5%, for a duration of 10 to 30 minutes. However, it is important to consider that Triton X-100 and NP-40 are relatively harsh detergents. When used at higher concentrations or for prolonged incubation times, they may lead to the loss of soluble cytoplasmic and membrane proteins, perturbation of cellular structure, and potential alterations in antigen distribution and cellular integrity. #NonIonicDetergents #LipidSolubilization #PlasmaMembrane #Concentration #Detergents #Versatility #AntigenTargeting #StructuralIntegrity #AntigenDistribution

Tips and Tricks for Cell-Based Imaging Assays: Immunocytochemistry Fixation-Review The choice of fixative depends on several factors, including the nature of the antigen, the desired cellular morphology, and the experimental requirements. PFA is often preferred when preserving cellular morphology is crucial, especially for tissues with intricate structures. Methanol and ethanol are often preferred for detecting intracellular antigens and phosphoproteins due to their ability to rapidly permeabilize cells and preserve antigenicity. Acetone is commonly used for frozen sections and is effective in preserving delicate structures and maintaining antigen integrity. When fixing cells with organic solvents, it is advisable to wash the cells once or twice with phosphate-buffered saline (PBS) before applying the fixative. This eliminates traces of cell-culture media as organic solvents precipitate serum proteins in the media. It is important to note that fixative preference can also be influenced by the specific experimental setup and the compatibility of the fixative with subsequent ICC steps, such as permeabilization and antibody binding. Optimization of fixation conditions, including concentration, temperature, and duration, is necessary to ensure proper antigen preservation and minimize artifacts. #AntigenNature #OptimalPreservation #Detection #CellMorphology #Preservation #Structure #ExperimentalRequirements #Permeabilization #ProteinDetection