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Tracking and quantifying polymer therapeutic distribution on a cellular level using 3D dSTORM.

Jonathan M Hartley1, Rui Zhang2, Manasa Gudheti3

  • 1Department of Bioengineering, University of Utah, 20 S. 2030 E., Rm. 108, Salt Lake City, UT 84112, USA.

Journal of Controlled Release : Official Journal of the Controlled Release Society
|February 9, 2016
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Summary

Direct stochastic optical reconstruction microscopy (dSTORM) visualized nanoscale drug delivery systems. This technique quantified nanoconjugate distribution and colocalization, revealing insights into therapeutic component behavior within cells.

Keywords:
Drug deliveryN-(2-hydroxypropyl)methacrylamide (HPMA)Super resolutiondSTORM

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Area of Science:

  • Biotechnology
  • Nanomedicine
  • Cell Biology

Background:

  • N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer conjugates are advanced drug delivery systems.
  • Understanding the nanoscale distribution and colocalization of these therapeutics is crucial for optimizing their efficacy.
  • Existing imaging techniques often lack the resolution to visualize these processes at the cellular level.

Purpose of the Study:

  • To apply direct stochastic optical reconstruction microscopy (dSTORM) for high-resolution imaging of HPMA copolymer conjugates.
  • To quantify the colocalization and spatial distribution of therapeutic components within cells.
  • To demonstrate the versatility of dSTORM for studying various drug delivery systems.

Main Methods:

  • Utilized direct stochastic optical reconstruction microscopy (dSTORM) for nanoscale imaging.
  • Acquired high-resolution (25nm) microscopy images of entire cells.
  • Employed spatial descriptive statistics to analyze drug and polymer distribution.

Main Results:

  • dSTORM revealed nanoscale distribution of fluorescently labeled therapeutic components.
  • Drug-free nanoconjugates showed increased nanoclusters on cell surfaces over time.
  • An ovarian cancer conjugate demonstrated drug-polymer colocalization before enzymatic cleavage, followed by random drug distribution and polymer clustering at 24h.

Conclusions:

  • dSTORM is a versatile technique for visualizing and quantifying the nanoscale behavior of drug delivery systems.
  • The study provides insights into the spatial dynamics of HPMA copolymer conjugates within cellular environments.
  • dSTORM's high resolution is valuable for understanding therapeutic component interactions and drug release mechanisms.