Biomod/2013/OSU/protocol/cell analysis

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Cell Analysis Procedure

Loading Efficiency

  1. Setup a 2 mM Daunorubicin Solution (Given Concentration of Solution)
  2. Add Origami shapes at a concentration of 5.0 nM
  3. Incubate for 40 hours
  4. Centrifuge at 20.0 xG for 50 minutes @ 20 deg C
  5. Isolate Orange-red precipitate from solution - PEG (Keep solution—gives concentration of daunorubicin not absorbed by origami/used in loading efficiency calculation)
  6. Read Absorption at 488 nm with a microplate reader (FRET Setting)

Cellular Methods

  1. Redissolve origami shapes in PBS to form Stock solution (~1 mM Daunorubicin and~1.25 nM origami)
  2. Dilute with cell culture medium to form final Daunorubicin concentrations of 5 to 100 µm

Cellular Viability (MTT)

  1. Seed 96-Well plate with CLL cells and incubate overnight
  2. Add Origami Shapes, double stranded DNA, or free doxorubicin at specified concentration to cells for 12 hours
  3. Wash with PBS
  4. Incubate for 24, 36, 48 hours
  5. Incubate with fresh serum-free medium containing 0.5 mg/mL WST-1 for 1 h at 37 °C for the cytotoxicity assay (Specific to cell-counting kit used)
  6. Read absorbance at 450 nm using microplate reader

Dead Cell Staining (TIRF)

  1. Seed 8-Well plate with CLL cells and incubate overnight
  2. Add Origami shapes, double stranded DNA, or free doxorubicin at specified concentration to cells for 12 hours
  3. Wash with PBS
  4. Incubate for 48 hours
  5. Incubate with SYTOX green (10 µM) for 20 minutes at 37ºC
  6. Image using Fluorescence Microscopy


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