6. Analyze the degree of scarring of individual “wounds” at
subsequent time points using wound-healing assays analysis
software.
4 Notes
1. Bubbling N 2 at for 20 min in order to prevent unwanted
oxidation of nanoparticles during the synthesis. Remember to
keep the mixture all the time under N 2 atmosphere during the
synthesis.
2. The collection of nanoparticles by external magnet can be
performed in the beaker rather than in round bottom flask.
The flat bottom of beaker gives bigger surface to collect nanoparticles and allows to handle them during the washing steps.
3. All activities related to cell culture perform under sterile conditions underneath a laminar flow cabinet.
4. Images of the Mag@PEI NPs were visualized using 559 nm
excitation and 570–590 nm emission filters. To visualize the
cell membranes, 488 nm excitation and 495–545 nm emission
filters were applied. The Hoechst fluorescence was detected
using 405 nm excitation source and 425–475 nm emission
filters.
5. Images of the Mag@PEI NPs were visualized using 559 nm
excitation and 570–590 nm emission filters, whereas
ATN-RNA was visualized using 488 nm excitation and
495–545 nm emission filters. The Hoechst fluorescence was
detected using 405 nm excitation source and 425–475 nm
emission filters.
6. Perform all operations involving RNA manipulations on ice,
using nuclease-free water.
7. Using a real-time PCR system based on probes allows for
greater specificity of detection than a system based on
fluorescent dyes.
Acknowledgments
The financial support under project number 2016/21/B/ST8/
00477 granted by The National Science Centre, Poland to S.Jurga
is kindly acknowledged.
80
Małgorzata Grabowska et al.
subsequent time points using wound-healing assays analysis
software.
4 Notes
1. Bubbling N 2 at for 20 min in order to prevent unwanted
oxidation of nanoparticles during the synthesis. Remember to
keep the mixture all the time under N 2 atmosphere during the
synthesis.
2. The collection of nanoparticles by external magnet can be
performed in the beaker rather than in round bottom flask.
The flat bottom of beaker gives bigger surface to collect nanoparticles and allows to handle them during the washing steps.
3. All activities related to cell culture perform under sterile conditions underneath a laminar flow cabinet.
4. Images of the Mag@PEI NPs were visualized using 559 nm
excitation and 570–590 nm emission filters. To visualize the
cell membranes, 488 nm excitation and 495–545 nm emission
filters were applied. The Hoechst fluorescence was detected
using 405 nm excitation source and 425–475 nm emission
filters.
5. Images of the Mag@PEI NPs were visualized using 559 nm
excitation and 570–590 nm emission filters, whereas
ATN-RNA was visualized using 488 nm excitation and
495–545 nm emission filters. The Hoechst fluorescence was
detected using 405 nm excitation source and 425–475 nm
emission filters.
6. Perform all operations involving RNA manipulations on ice,
using nuclease-free water.
7. Using a real-time PCR system based on probes allows for
greater specificity of detection than a system based on
fluorescent dyes.
Acknowledgments
The financial support under project number 2016/21/B/ST8/
00477 granted by The National Science Centre, Poland to S.Jurga
is kindly acknowledged.
80
Małgorzata Grabowska et al.