3D Spheroid & Organoid System:

  • Generation of 3D cancer spheroids and organoids
  • Drug screening and drug toxicity assays
  • Whole spheroids/ organoids staining & imaging

Cell Viability Assays:

  • MTT/MTS assays: Measure cell metabolic activity through the reduction of tetrazolium compounds.
  • CellTiter-Glo®: It is a cell assay that measures cell viability by quantifying ATP released from living cells, providing a direct indication of the number of viable cells in a culture.
  • Resazurin reduction assay (Alamar Blue): Detect metabolic activity through resazurin reduction.

Cytotoxicity Assays:

  • Lactate dehydrogenase (LDH) release assay: Detects cell membrane damage by measuring LDH release.
  • Trypan blue exclusion assay: Counts viable cells that exclude the dye.
  • Propidium iodide staining: Identifies dead cells by staining DNA.

Apoptosis Assays:

  • Caspase 3/7 Assay: Measures the activation of 3/7 caspases which are key enzymes that execute apoptosis, or programmed cell death in cells.
  • Annexin V staining: Detects phosphatidylserine exposure on the outer leaflet of the plasma membrane.
  • TUNEL assay (Terminal deoxynucleotidyl transferase dUTP nick end labelling): Identifies DNA fragmentation.

Proliferation Assays:

  • BrdU (Bromodeoxyuridine) incorporation assay: Measures DNA synthesis by incorporating BrdU into newly synthesized DNA.
  • Ki-67 staining: Detects the proliferation marker Ki-67 in cells.
  • EdU (5-ethynyl-2'-deoxyuridine) incorporation assay: A more modern alternative to BrdU.
  • Cell cycle analysis: Cell cycle analysis by DNA content measurement is commonly performed using flow cytometry with fluorescent dyes like propidium iodide or DAPI. The fluorescence intensity correlates with the amount of DNA in cells, allowing differentiation of cells in different phases of the cell cycle.

Signal Transduction Assays:

  • Calcium flux assays: Measure intracellular calcium levels using fluorescent indicators.
  • cAMP assays: Quantify cyclic AMP levels as a marker of GPCR activation.
  • Phosphorylation assays: Detect the phosphorylation status of proteins using antibodies.

Selecting Fashion Theme

  • Luminex assays: Simultaneously measure multiple analytes in a single sample using bead-based technology.
  • Cytokine profiling: Quantify multiple cytokines secreted by cells.

Enzymatic assays:

  • Reactive Oxygen Species (ROS): The fluorescent dyes like DCFDA that react with ROS to produce a fluorescent signal is used. The cells are incubated with the probe, stimulated to produce ROS, and the fluorescence is measured.
  • Nitric Oxide Synthase (NOS): The NOS assay is crucial in understanding various biological processes, such as neurotransmission, vascular regulation, immune response, and apoptosis, where NO plays a significant role. The assay involves a series of reactions that convert NO into nitrite and nitrate, which are then quantified using colorimetric or spectrophotometric methods.

Cell Migration and Invasion Assays:

  • Scratch (wound healing) assay: Measures cell migration into a scratched area on a cell monolayer.
  • Migration assay: Assesses the movement of cells through a membrane towards a chemoattractant.
  • Matrigel invasion assay: Evaluates cell invasion through a Matrigel-coated membrane.

High-content Screening (HCS):

  • Automated microscopy: Captures and analyses multiple cellular parameters.
  • Image-based assays: Quantify cellular features such as morphology, protein localization, and cell cycle.

Cell Differentiation Assays:

  • Osteogenesis, adipogenesis, and myogenesis assays: Measure the differentiation of stem cells into specific lineages.
  • Neurite outgrowth assays: Assess the differentiation and growth of neurons.

Permeability Assay:

  • CaCO2: It is a gold standard method for evaluating both passive and active transport and absorption of orally administered drugs, making it an indispensable tool in drug development
  • Parallel Artificial Membrane Permeability Assay (PAMPA): It is a complementary tool to evaluate the passive transportation of drugs.

Receptor Internalization and Trafficking Assays:

  • Fluorescent ligand binding: Visualize receptor internalization using labelled ligands.
  • Tag-based assays (e.g., HA, GFP-tagged receptors): Monitor receptor trafficking.