June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Voltage-coupled single-needle constant-pressure anterior chamber perfusion in live mice
Author Affiliations & Notes
  • MinHee Ko
    Ophthalmology, University of Southern California, Los Angeles, CA
  • Aleksandr Yelenskiy
    School of Medicine, University of Wisconsin, Madison, WI
  • Jose Gonzalez
    Ophthalmology, University of Southern California, Los Angeles, CA
  • James Tan
    Ophthalmology, University of Southern California, Los Angeles, CA
  • Footnotes
    Commercial Relationships MinHee Ko, None; Aleksandr Yelenskiy, None; Jose Gonzalez, None; James Tan, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2001. doi:
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      MinHee Ko, Aleksandr Yelenskiy, Jose Gonzalez, James Tan; Voltage-coupled single-needle constant-pressure anterior chamber perfusion in live mice. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2001.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: To describe voltage-coupled single-needle constant-pressure anterior chamber perfusion in live mice

Methods: We established apparatus in which transduced pressure was coupled to a microperfusion pump to automatically modulate flow rate to maintain a steady pressure and measure outflow facility by constant pressure perfusion. All perfusion was performed by 35G single-needle anterior chamber cannulation in anesthetized mice. We characterized the following: (i) perfusion pressure stability; (ii) outflow facility; (iii) presence of “washout”; (iv) effect of different perfusates such as phosphate buffered saline with Ca2+and Mg2+ (PBS w Ca/Mg), PBS without Ca/Mg (PBS w/o Ca/Mg), and Barany’s solution. H&E staining was performed on frozen sections following perfusions.

Results: Twenty nine live C57BL/6 mice underwent perfusion. Constant pressure was attained within seconds, stably maintained, and not significantly affected by different perfusates (p>0.05). Relationship between flow rate and pressure fit a linear function for perfusion between 15-35mmHg (R2>0.9). Outflow facility determined by 1-level constant pressure perfusion was similar irrespective of perfusate (p>0.5): 0.0095 μl/min*mmHg for PBS w Ca/Mg (n=10); 0.0123 μl/min*mmHg for PBS w/o Ca/Mg (n=10); 0.0074 μl/min*mmHg for Barany’s solution (n=9). Needle resistance was negligible relative to physiologically relevant perfusion. 2-level constant pressure perfusion over 2 hours showed no washout phenomenon. Histological disruption of drainage tissue following perfusions was not seen.

Conclusions: Stable constant pressure perfusion under physiologically relevant conditions was achieved by perfusion using single-needle cannulation, which is well-suited to the tiny mouse eye. The washout phenomenon was not seen. Our methods are potentially applicable to live mouse studies.

Keywords: 633 outflow: trabecular meshwork • 735 trabecular meshwork  
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